DOCSLIB.ORG

Characterization of the Unique Flagellar Hook Structure of the Spirochetes Borrelia Burgdorferi and Treponema Denticola

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text

Load more

Graduate Theses, Dissertations, and Problem Reports 2013 Characterization of the Unique Flagellar Hook Structure of the Spirochetes Borrelia burgdorferi and Treponema denticola Kelly Ann Heintzelman Miller West Virginia University Follow this and additional works at: https://researchrepository.wvu.edu/etd Recommended Citation Miller, Kelly Ann Heintzelman, "Characterization of the Unique Flagellar Hook Structure of the Spirochetes Borrelia burgdorferi and Treponema denticola" (2013). Graduate Theses, Dissertations, and Problem Reports. 4983. https://researchrepository.wvu.edu/etd/4983 This Dissertation is protected by copyright and/or related rights. It has been brought to you by the The Research Repository @ WVU with permission from the rights-holder(s). You are free to use this Dissertation in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you must obtain permission from the rights-holder(s) directly, unless additional rights are indicated by a Creative Commons license in the record and/ or on the work itself. This Dissertation has been accepted for inclusion in WVU Graduate Theses, Dissertations, and Problem Reports collection by an authorized administrator of The Research Repository @ WVU. For more information, please contact [email protected]. Characterization of the Unique Flagellar Hook Structure of the Spirochetes Borrelia burgdorferi and Treponema denticola Kelly Ann Heintzelman Miller Dissertation submitted to the School of Medicine at West Virginia University in partial fullfillment of the requirements for the degree of Doctor of Philosophy in Immunology and Microbial Pathogenesis Nyles W. Charon, Ph.D., Chair Michael R. Miller, Ph.D. Thomas Elliott, Ph.D. Slawomir Lukomski, Ph.D. Rita Rio, Ph.D. Department of Microbial Pathogenesis and Immunology Morgantown, WV 2013 Keywords: flagella, hook, Borrelia burgdorferi, Treponema denticola, mass spectrometry, peptide cross-linking, spirochete ABSTRACT Characterization of the Unique Flagellar Hook Structure of the Spirochetes Borrelia burgdorferi and Treponema denticola Kelly Ann Heintzelman Miller The long-term goal of this project is to better understand Borrelia burgdorferi and Treponema denticola motility as it relates to pathogenesis. The structure of the spirochete periplasmic flagella is unique when compared to externally located flagella. One unusual characteristic is the flagellar hook, which serves as a universal joint coupling rotation of the membrane bound motor complex to the flagellar filament. The hook is composed of approximately 120 FlgE monomers, and in bacteria with externally located flagella, these structures readily disassociate into monomers (~50 kDa) when treated with heat and detergent. However, in spirochetes the FlgE monomers form a large mass of over 250 kDa. We hypothesize that this high molecular weight complex (HMWC) is the result of covalent cross-linking of FlgE proteins. Cross-linking of bacterial flagella proteins has not been examined previously. Our goals are to thoroughly characterize the cross-linking in the B.burgdorferi and T. denticola hook proteins. We anticipate that once we have this information, new drugs could be developed to inhibit this cross-linking, which we anticipate will severely affect the spirochetes' motility and virulence capability. Thus, the proposed project’s long-term goal is centered on developing new drug treatments for Lyme disease as well as other spirochetal diseases, including syphilis. We found that the HMWC of both B. burgdorferi and T. denticola are stable to a variety of denaturants and is not an artifact of boiling. Previously, western blot analysis was the sole indication that spirochetes cross- link their flagellar hook proteins. By making several modifications to the published procedure for isolating periplasmic flagella, I was able to obtain enough hook protein of both B. burgdorferi and T. denticola for mass spectrometry analysis. The mass spectrometry analysis confirmed that the spirochete HMWC is composed of FlgE, identified FlgE as the sole protein in these complexes, and that in specific regions the mass spectrometry patterns are different for the monomer and HMWC. We observed that T. denticola FlgE over-expressed in Escherichia coli forms a stable HMWC in vitro, and used the in vitro HMWC to study the effects of both chemical treatments and point mutations on HMWC formation. Finally, I propose a mechanism of cross-linking that is consistent with the mass spectrometry, biochemical, and mutational analyses. We hypothesize that FlgE cross-linking strengthens the hook structure for optimal spirochete motility. Acknowledgements I wish to dedicate this work to my snuggle buddy Kodiak Lee. You are the joy that keeps me going over day through the highs and lows. You have taught me patience and make me laugh when I'm about to cry. There are several people I would like to thank for support and guidance through this journey. Thank God. Without him, nothing is possible. My husband Eric for patience, listening to me vent, and comforting me when stressed. My parents for their support, time, and taking care of Kodiak. Milinda for being a rock and maintaining my sanity. Giving me support both inside and outside of lab. My mentors Nyles and Mike for taking on one more graduate student before retirement, letting me work on a really cool project, guiding me through this process, and for being good friends. My friends Amanda, Dacie, and Chrissy. For always being there, for good times, and helping me with the job search. Andy Cockburn for listening and good suggestions. You always looked at things from a different angle and gave fresh ideas. My committee: Thomas Elliott, Slawomir Lukomski, and Rita Rio for ideas, suggestions, looking over results with me, assisting in experimental design, and facilitating my scientific development. iii Collaborators: Chris Li (SUNY, Buffalo), Brian Crane (Cornell), Jun Liu (University of Texas), MD Motaleb (ECU), Justin Hettick (NIOSH), Melissa Caimano (University of Connecticut), Roger Hendrix and Bob Duda (University of Pittsburgh). Thank you for your contributions to this project. My dogs Lucky and Reba for stress relief and nice walks. iv Table of Contents Chapter 1: Introduction....................................................................................... 1 Medical Importance of T. denticola................................................................... 2 Medical Importance of B. burgdorferi............................................................... 3 Flagellar structure and motility.......................................................................... 4 Structure of periplasmic flagella and how spirochetes swim............................ 12 Motility as a virulence factor of spirochetes...................................................... 15 Covalent bonding of peptides in microorganisms............................................. 17 Methodology to detecting and deciphering cross-links..................................... 22 Indirect evidence of cross-linking in spirochete flagella.................................... 23 Research objectives......................................................................................... 25 References............................................................................................... 26 Chapter 2: Initial characterization of the FlgE high molecular weight complex of Borrelia burgdorferi......................................................................... 39 Abstract............................................................................................................ 40 Introduction....................................................................................................... 41 Materials and Methods..................................................................................... 43 Results and Discussion.................................................................................... 48 References............................................................................................... 59 Chapter 3: The biochemistry of the Treponema denticola FlgE high molecular weight complex.................................................................................. 68 Abstract............................................................................................................ 69 Introduction....................................................................................................... 70 Materials and Methods..................................................................................... 71 Results.............................................................................................................. 76 Discussion........................................................................................................ 98 References....................................................................................................... 104 Chapter 4: Discussion......................................................................................... 111 Appendix 122 The unique paradigm of spirochete motility and chemotaxis............................ 130 Visualizing the sequential assembly of bacterial flagella in Borrelia 154 burgdorferi by cryo-electron tomography.......................................................... Curriculum Vitae 176 v List of Figures Figure 1.1: Flagellar components of S. enterica...................................... 4 Figure 1.2: Proposed structure
Recommended publications
  • LYME DISEASE Other Names: Borrelia Burgdorferi

    LYME DISEASE Other Names: Borrelia Burgdorferi

    LYME DISEASE Other names: Borrelia burgdorferi CAUSE Lyme disease is caused by a spirochete bacteria (Borrelia burgdorferi) that is transmitted through the bite from an infected arthropod vector, the black-legged or deer tick Ixodes( scapularis). SIGNIFICANCE Lyme disease can infect people and some species of domestic animals (cats, dogs, horses, and cattle) causing mild to severe illness. Although wildlife can be infected by the bacteria, it typically does not cause illness in them. TRANSMISSION The bacteria has been observed in the blood of a number of wildlife species including several bird species but rarely appears to cause illness in these species. White-footed mice, eastern chipmunks, and shrews serve as the primary natural reservoirs for Lyme disease in eastern and central parts of North America. Other species appear to have low competencies as reservoirs for the bacteria. The transmission of Lyme disease is relatively convoluted due to the complex life cycle of the black-legged tick. This tick has multiple developmental stages and requires three hosts during its life cycle. The life cycle begins with the eggs of the ticks that are laid in the spring and from which larval ticks emerge. Larval ticks do not initially carryBorrelia burgdorferi, the bacteria must be acquired from their hosts they feed upon that are carriers of the bacteria. Through the summer the larval ticks feed on the blood of their first host, typically small mammals and birds. It is at this point where ticks may first acquireBorrelia burgdorferi. In the fall the larval ticks develop into nymphs and hibernate through the winter.
  • Differences in Genotype, Clinical Features, and Inflammatory

    Differences in Genotype, Clinical Features, and Inflammatory

    RESEARCH Differences in Genotype, Clinical Features, and Inflammatory Potential of Borrelia burgdorferi sensu stricto Strains from Europe and the United States Tjasa Cerar,1 Franc Strle,1 Dasa Stupica, Eva Ruzic-Sabljic, Gail McHugh, Allen C. Steere, Klemen Strle Borrelia burgdorferi sensu stricto isolates from patients with B. afzelii, which usually remains localized to the skin, with erythema migrans in Europe and the United States and B. garinii, which is usually associated with nervous were compared by genotype, clinical features of infection, system involvement (1). B. burgdorferi infection is rare in and inflammatory potential. Analysis of outer surface pro- Europe; little is known about its clinical course there. In tein C and multilocus sequence typing showed that strains the United States, B. burgdorferi is the sole agent of Lyme from these 2 regions represent distinct genotypes. Clinical borreliosis; in the northeastern United States, it is particu- features of infection with B. burgdorferi in Slovenia were similar to infection with B. afzelii or B. garinii, the other 2 larly arthritogenic (1,2). For all 3 species, the first sign of Borrelia spp. that cause disease in Europe, whereas B. infection is often an erythema migrans lesion. However, B. burgdorferi strains from the United States were associ- burgdorferi infection in the United States is associated with ated with more severe disease. Moreover, B. burgdorferi a greater number of disease-associated symptoms and more strains from the United States induced peripheral blood frequent hematogenous dissemination than B. afzelii or B. mononuclear cells to secrete higher levels of cytokines garinii infection in Europe (4–7).
  • Canine Lyme Borrelia

    Canine Lyme Borrelia

    Canine Lyme Borrelia Borrelia burgdorferi bacteria are the cause of Lyme disease in humans and animals. They can be visualized by darkfild microscopy as "corkscrew-shaped" motile spirochetes (400 x). Inset: The black-legged tick, lxodes scapularis (deer tick), may carry and transmit Borrelia burgdorferi to humans and animals during feeding, and thus transmit Lyme disease. Samples: Blood EDTA-blood as is, purple-top tubes or EDTA-blood preserved in sample buffer (preferred) Body fluids Preserved in sample buffer Notes: Send all samples at room temperature, preferably preserved in sample buffer MD Submission Form Interpretation of PCR Results: High Positive Borrelia spp. infection (interpretation must be correlated to (> 500 copies/ml swab) clinical symptoms) Low Positive (<500 copies/ml swab) Negative Borrelia spp. not detected Lyme Borreliosis Lyme disease is caused by spirochete bacteria of a subgroup of Borrelia species, called Borrelia burgdorferi sensu lato. Only one species, B. burgdorferi sensu stricto, is known to be present in the USA, while at least four pathogenic species, B. burgdorferi sensu stricto, B. afzelii, B. garinii, B. japonica have been isolated in Europe and Asia (Aguero- Rosenfeld et al., 2005). B. burgdorferi sensu lato organisms are corkscrew-shaped, motile, microaerophilic bacteria of the order Spirochaetales. Hard-shelled ticks of the genus Ixodes transmit B. burgdorferi by attaching and feeding on various mammalian, avian, and reptilian hosts. In the northeastern states of the US Ixodes scapularis, the black-legged deer tick, is the predominant vector, while at the west coast Lyme borreliosis is maintained by a transmission cycle which involves two tick species, I.
  • Borrelia Burgdorferi and Treponema Pallidum: a Comparison of Functional Genomics, Environmental Adaptations, and Pathogenic Mechanisms

    Borrelia Burgdorferi and Treponema Pallidum: a Comparison of Functional Genomics, Environmental Adaptations, and Pathogenic Mechanisms

    PERSPECTIVE SERIES Bacterial polymorphisms Martin J. Blaser and James M. Musser, Series Editors Borrelia burgdorferi and Treponema pallidum: a comparison of functional genomics, environmental adaptations, and pathogenic mechanisms Stephen F. Porcella and Tom G. Schwan Laboratory of Human Bacterial Pathogenesis, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, NIH, Hamilton, Montana, USA Address correspondence to: Tom G. Schwan, Rocky Mountain Laboratories, 903 South 4th Street, Hamilton, Montana 59840, USA. Phone: (406) 363-9250; Fax: (406) 363-9445; E-mail: [email protected]. Spirochetes are a diverse group of bacteria found in (6–8). Here, we compare the biology and genomes of soil, deep in marine sediments, commensal in the gut these two spirochetal pathogens with reference to their of termites and other arthropods, or obligate parasites different host associations and modes of transmission. of vertebrates. Two pathogenic spirochetes that are the focus of this perspective are Borrelia burgdorferi sensu Genomic structure lato, a causative agent of Lyme disease, and Treponema A striking difference between B. burgdorferi and T. pal- pallidum subspecies pallidum, the agent of venereal lidum is their total genomic structure. Although both syphilis. Although these organisms are bound togeth- pathogens have small genomes, compared with many er by ancient ancestry and similar morphology (Figure well known bacteria such as Escherichia coli and Mycobac- 1), as well as by the protean nature of the infections terium tuberculosis, the genomic structure of B. burgdorferi they cause, many differences exist in their life cycles, environmental adaptations, and impact on human health and behavior. The specific mechanisms con- tributing to multisystem disease and persistent, long- term infections caused by both organisms in spite of significant immune responses are not yet understood.
  • Investigation of the Lipoproteome of the Lyme Disease Bacterium

    Investigation of the Lipoproteome of the Lyme Disease Bacterium

    INVESTIGATION OF THE LIPOPROTEOME OF THE LYME DISEASE BACTERIUM BORRELIA BURGDORFERI BY Alexander S. Dowdell Submitted to the graduate degree program in Microbiology, Molecular Genetics & Immunology and the Graduate Faculty of the University of Kansas in partial fulfillment of the requirements for the degree of Doctor of Philosophy. _____________________________ Wolfram R. Zückert, Ph.D., Chairperson _____________________________ Indranil Biswas, Ph.D. _____________________________ Mark Fisher, Ph.D. _____________________________ Joe Lutkenhaus, Ph.D. _____________________________ Michael Parmely, Ph.D. Date Defended: April 27th, 2017 The dissertation committee for Alexander S. Dowdell certifies that this is the approved version of the following dissertation: INVESTIGATION OF THE LIPOPROTEOME OF THE LYME DISEASE BACTERIUM BORRELIA BURGDORFERI _____________________________ Wolfram R. Zückert, Ph.D., Chairperson Date Approved: May 4th, 2017 ii Abstract The spirochete bacterium Borrelia burgdorferi is the causative agent of Lyme borreliosis, the top vector-borne disease in the United States. B. burgdorferi is transmitted by hard- bodied Ixodes ticks in an enzootic tick/vertebrate cycle, with human infection occurring in an accidental, “dead-end” fashion. Despite the estimated 300,000 cases that occur each year, no FDA-approved vaccine is available for the prevention of Lyme borreliosis in humans. Development of new prophylaxes is constrained by the limited understanding of the pathobiology of B. burgdorferi, as past investigations have focused intensely on just a handful of identified proteins that play key roles in the tick/vertebrate infection cycle. As such, identification of novel B. burgdorferi virulence factors is needed in order to expedite the discovery of new anti-Lyme therapeutics. The multitude of lipoproteins expressed by the spirochete fall into one such category of virulence factor that merits further study.
  • Information to Users

    Information to Users

    INFORMATION TO USERS This manuscript bas been reproJuced from the microfilm master. UMI films the text directly ftom the original or copy submitted. Thus, sorne thesis and dissertation copies are in typewriter face, while others may be itom any type ofcomputer printer. The quality oftbis reproduction is depeDdeDt apoD the quality of the copy sablDitted. Broken or indistinct print, colored or poor quality illustrations and photographs, print bleedthlough, substandard margins, and improper alignment can adversely affect reproduction. In the unlikely event that the author did not send UMI a complete manuscript and there are missing pages, these will he noted. Also, if unauthorized copyright material had to be removed, a note will indicate the deletion. Oversize materials (e.g., maps, drawings, charts) are reproduced by sectioning the original, beginning at the upper left-hand corner and continuing trom left to right in equal sections with sma1l overlaps. Each original is a1so photographed in one exposure and is included in reduced fonn at the back orthe book. Photographs ineluded in the original manuscript have been reproduced xerographically in this copy. Higher quality 6" x 9" black and white photographie prints are available for any photographs or illustrations appearing in this copy for an additional charge. Contact UMI directly to order. UMI A Bell & Howell Information Company 300 North Zeeb Raad, ADn AJbor MI 48106-1346 USA 313n61-4700 8OO1S21~ NOTE TO USERS The original manuscript received by UMI contains pages with slanted print. Pages were microfilmed as received. This reproduction is the best copy available UMI Oral spirochetes: contribution to oral malodor and formation ofspherical bodies by Angela De Ciccio A thesis submitted to the Faculty ofGraduate Studies and Research, McGill University, in partial fulfillment ofthe requirements for the degree ofMaster ofScience.
  • Detection of Putative Periodontal Pathogens in Subgingival Specimens of Dogs

    Detection of Putative Periodontal Pathogens in Subgingival Specimens of Dogs

    Brazilian Journal of Microbiology (2007) 38:23-28 ISSN 1517-8283 DETECTION OF PUTATIVE PERIODONTAL PATHOGENS IN SUBGINGIVAL SPECIMENS OF DOGS Sheila Alexandra Belini Nishiyama1; Gerusa Neyla Andrade Senhorinho1; Marco Antônio Gioso2; Mario Julio Avila-Campos1,* 1Anaerobe Laboratory, Department of Microbiology, Institute of Biomedical Science; 2Veterinary and Zootechny School, University of São Paulo, São Paulo, SP, Brazil Submitted: April 07, 2006; Returned to authors for corrections: July 13, 2006; Approved: October 13, 2006 ABSTRACT In this study, the presence of putative periodontal organisms, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythensis, Fusobacterium nucleatum, Dialister pneumosintes, Actinobacillus actinomycetemcomitans, Campylobacter rectus, Eikenella corrodens and Treponema denticola were examined from subgingival samples of 40 dogs of different breeds with (25) and without (15) periodontitis, by using the PCR method. The PCR products of each species showed specific amplicons. Of the 25 dogs with periodontitis, P. gingivalis was detected in 16 (64%) samples, C. rectus in 9 (36%), A. actinomycetemcomitans in 6 (24%), P. intermedia in 5 (20%), T. forsythensis in 5 (20%), F. nucleatum in 4 (16%) and E. corrodens in 3 (12%). T. denticola and D. pneumosintes were not detected in clinical samples from dogs with periodontitis. Moreover, P. gingivalis was detected only in one (6.66%) crossbred dog without periodontitis. Our results show that these microorganisms are present in periodontal microbiota of dogs with periodontitits, and it is important to evaluate the role of these putative periodontal microorganisms play in the periodontitis in household pets particularly, dogs in ecologic and therapeutic terms, since these animals might acquire these periodontopahogens from their respective owners.
  • Taxonomy JN869023

    Taxonomy JN869023

    Species that differentiate periods of high vs. low species richness in unattached communities Species Taxonomy JN869023 Bacteria; Actinobacteria; Actinobacteria; Actinomycetales; ACK-M1 JN674641 Bacteria; Bacteroidetes; [Saprospirae]; [Saprospirales]; Chitinophagaceae; Sediminibacterium JN869030 Bacteria; Actinobacteria; Actinobacteria; Actinomycetales; ACK-M1 U51104 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Comamonadaceae; Limnohabitans JN868812 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Comamonadaceae JN391888 Bacteria; Planctomycetes; Planctomycetia; Planctomycetales; Planctomycetaceae; Planctomyces HM856408 Bacteria; Planctomycetes; Phycisphaerae; Phycisphaerales GQ347385 Bacteria; Verrucomicrobia; [Methylacidiphilae]; Methylacidiphilales; LD19 GU305856 Bacteria; Proteobacteria; Alphaproteobacteria; Rickettsiales; Pelagibacteraceae GQ340302 Bacteria; Actinobacteria; Actinobacteria; Actinomycetales JN869125 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Comamonadaceae New.ReferenceOTU470 Bacteria; Cyanobacteria; ML635J-21 JN679119 Bacteria; Proteobacteria; Betaproteobacteria; Burkholderiales; Comamonadaceae HM141858 Bacteria; Acidobacteria; Holophagae; Holophagales; Holophagaceae; Geothrix FQ659340 Bacteria; Verrucomicrobia; [Pedosphaerae]; [Pedosphaerales]; auto67_4W AY133074 Bacteria; Elusimicrobia; Elusimicrobia; Elusimicrobiales FJ800541 Bacteria; Verrucomicrobia; [Pedosphaerae]; [Pedosphaerales]; R4-41B JQ346769 Bacteria; Acidobacteria; [Chloracidobacteria]; RB41; Ellin6075
  • Assessing Lyme Disease Relevant Antibiotics Through Gut Bacteroides Panels

    Assessing Lyme Disease Relevant Antibiotics Through Gut Bacteroides Panels

    Assessing Lyme Disease Relevant Antibiotics through Gut Bacteroides Panels by Sohum Sheth Abstract: Lyme borreliosis is the most prevalent vector-borne disease in the United States caused by the transmission of bacteria Borrelia burgdorferi harbored by the Ixodus scapularis ticks (Sharma, Brown, Matluck, Hu, & Lewis, 2015). Antibiotics currently used to treat Lyme disease include oral doxycycline, amoxicillin, and ce!riaxone. Although the current treatment is e"ective in most cases, there is need for the development of new antibiotics against Lyme disease, as the treatment does not work in 10-20% of the population for unknown reasons (X. Wu et al., 2018). Use of antibiotics in the treatment of various diseases such as Lyme disease is essential; however, the downside is the development of resistance and possibly deleterious e"ects on the human gut microbiota composition. Like other organs in the body, gut microbiota play an essential role in the health and disease state of the body (Ianiro, Tilg, & Gasbarrini, 2016). Of importance in the microbiome is the genus Bacteroides, which accounts for roughly one-third of gut microbiome composition (H. M. Wexler, 2007). $e purpose of this study is to investigate how antibiotics currently used for the treatment of Lyme disease in%uences the Bacteroides cultures in vitro and compare it with a new antibiotic (antibiotic X) identi&ed in the laboratory to be e"ective against B. burgdorferi. Using microdilution broth assay, minimum inhibitory concentration (MIC) was tested against nine di"erent strains of Bacteroides. Results showed that antibiotic X has a higher MIC against Bacteroides when compared to amoxicillin, ce!riaxone, and doxycycline, making it a promising new drug for further investigation and in vivo studies.
  • Molecular Studies of Treponema Pallidum

    Molecular Studies of Treponema Pallidum

    Fall 08 Molecular Studies of Treponema pallidum Craig Tipple Imperial College London Department of Medicine Section of Infectious Diseases Thesis submitted in fulfillment of the requirements for the degree of Doctor of Philosophy of Imperial College London 2013 1 Abstract Syphilis, caused by Treponema pallidum (T. pallidum), has re-emerged in the UK and globally. There are 11 million new cases annually. The WHO stated the urgent need for single-dose oral treatments for syphilis to replace penicillin injections. Azithromycin showed initial promise, but macrolide resistance-associated mutations are emerging. Response to treatment is monitored by serological assays that can take months to indicate treatment success, thus a new test for identifying treatment failure rapidly in future clinical trials is required. Molecular studies are key in syphilis research, as T. pallidum cannot be sustained in culture. The work presented in this thesis aimed to design and validate both a qPCR and a RT- qPCR to quantify T. pallidum in clinical samples and use these assays to characterise treatment responses to standard therapy by determining the rate of T. pallidum clearance from blood and ulcer exudates. Finally, using samples from three cross-sectional studies, it aimed to establish the prevalence of T. pallidum strains, including those with macrolide resistance in London and Colombo, Sri Lanka. The sensitivity of T. pallidum detection in ulcers was significantly higher than in blood samples, the likely result of higher bacterial loads in ulcers. RNA detection during primary and latent disease was more sensitive than DNA and higher RNA quantities were detected at all stages. Bacteraemic patients most often had secondary disease and HIV-1 infected patients had higher bacterial loads in primary chancres.
  • Treponema Borrelia Family: Leptospiraceae Genus: Leptospira Gr

    Treponema Borrelia Family: Leptospiraceae Genus: Leptospira Gr

    Bacteriology lecture no.12 Spirochetes 3rd class -The spirochetes: are a large ,heterogeneous group of spiral ,motile bacteria. Although, • there are at least eight genera in this family ,only the genera Treponema,Borrelia,and Leptospira which contain organism pathogenic for humans . -There are some reports of intestinal spirochetes ,that have been isolated from biopsy material ,these are Brachyspira pilosicoli,and Brachyspira aalborgi. *Objectives* Taxonomy Order: Spirochaetales Family: Spirochaetaceae Genus: Treponema Borrelia Family: Leptospiraceae Genus: Leptospira -Gram-negative spirochetes -Spirochete from Greek for “coiled hair "they are : *1*Extremely thin and can be very long *2* Motile by periplasmic flagella (axial fibrils or endoflagella) *3*Outer sheath encloses axial fibrils *4*Axial fibrils originate from insertion pores at both poles of cell 1 Bacteriology lecture no.12 Spirochetes 3rd class Spirochaetales Associated Human Diseases Treponema Main Treponema are: - T. pallidum subspecies pallidum - Syphilis: Venereal (sexual) disease 2 Bacteriology lecture no.12 Spirochetes 3rd class - T. pertenue - Yaws Non venereal - T. carateum - Pinta skin disease All three species are morphologically identical Characteristics of T.pallidum 1-They are long ,slender ,helically coiled ,spiral or cork –screw shaped bacilli. 2-T.pallidum has an outer sheath or glycosaminoglycan contain peptidoglycan and maintain the structural integrity of the organisms. 3-Endoflagella (axial filament ) are the flagella-like organelles in the periplasmic space encased by the outer membranes . 4-The endoflagella begin at each end of the organism and wind around it ,extending to and overlapping at the midpoint. 5- Inside the endoflagella is the inner membrane (cytoplasmic membrane)that provide osmotic stability and cover the protoplasmic cylinders .
  • Phylogenetic Foundation of Spirochetes

    Phylogenetic Foundation of Spirochetes

    J. Mol. Microbiol. Biotechnol. (2000) 2(4): 341-344. JMMBSpirochete Symposium Phylogeny on341 Spirochete Physiology Phylogenetic Foundation of Spirochetes Bruce J. Paster* and Floyd E. Dewhirst Spirochaetales that is divided into three families; namely the Spirochaetaceae, the Brachyspiraceae, and the Department of Molecular Genetics, The Forsyth Institute, Leptospiraceae. The phylogenetic relationships of 140 Fenway, Boston, Massachusetts 02115, USA representatives of each genus are shown in Figure 1. The Spirochaetaceae are separated into 6 genera— Borrelia, Brevinema, Cristispira, Spirochaeta, “Spironema”, Abstract and Treponema. New genera of termite spirochetes, such as Clevelandina, Diplocalyx, and Hollandina, have been The spirochetes are free-living or host-associated, described on the basis of differences in ultrastructural traits helical bacteria, some of which are pathogenic to man (Breznak, 1984). It has been suggested that they also and animal. Comparisons of 16S rRNA sequences belong in the family Spirochaetaceae, but no sequence demonstrate that the spirochetes represent a information is presently available to determine their monophyletic phylum within the bacteria. The phylogenetic position within the spirochetes. spirochetes are presently classified in the Class The Brachyspiraceae contain the genus Brachyspira Spirochaetes in the order Spirochetales and are (Serpulina). Due to the close phylogenetic relationship of divided into three major phylogenetic groupings, or B. aarlborgi to species characterized as Serpulina, it has families. The first family Spirochaetaceae contains been recommended that a single genus be justified. Thus, species of the genera Borrelia, Brevinema, Cristispira, Brachyspira takes precedence over Serpulina since the Spirochaeta, Spironema, and Treponema. The second former genus was listed first as a valid name (Hovind- family Brachyspiraceae contains the genus Hougen et al., 1983).