Streptococcus Pneumoniae Capsular Polysaccharide Is Linked to Peptidoglycan Via a Direct Glycosidic Bond to Β-D-N-Acetylglucosamine
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The Fine Structure of Diplococcus Pneumoniae
THE FINE STRUCTURE OF DIPLOCOCCUS PNEUMONIAE ALEXANDER TOMASZ, Ph.D., JAMES D. JAMIESON, M.D., and ELENA OTTOLENGtII, M.D. From The Rockefeller Institute and the New York University School of Medicine, New York ABSTRACT The fine structure of an unencapsulated strain of Diplococcus pneumoniae is described. A strik- ing feature of thcsc bacteria is an intracytoplasmic membrane system which appears to be an extension of septa of dividing bactcria. The possible function of these structures and their relationship to the plasma membrane and other types of intracytoplasmic membranes found in pncumococcus is discussed. INTRODUCTION Our main interest in the fine structure of Diplo- walls. Throughout this paper, such preparations will coccus pneumoniae stems from the fact that these be referred to as "spheroplasts." bacteria readily undergo genetic transformation. The bacteria were fixed and stained according to the method of Ryter and Kellenberger (4), embedded Prior to undertaking electron microscope studies in cross-linked methacrylate, and sectioned with a on this process, the fine structure of pneumococcal Porter-Blum mlcrotome using a diamond knife. The cells in thin sections was examined. During the sections were stained with lead according to the preliminary stage of these studies on a transform- method of Karnovsky (5) (method B) and were able strain, we observed some unique membranous examined in the RCA electron microscopes models structures which, to the best of our knowledge, 2B, 3F, or in the Siemens Elmiskop I. have not previously been described in bacteria. RESULTS MATERIALS AND METHODS The nuclear region of pneumococcus resembles that Unencapsulated strains of Diplocoecus pneumoniae R6, of other bacteria prepared by the method of R1, and some nutritional mutants derived from R6 Ryter and Kellenberger (4). -
Infection at the Wildlife-Livestock-Human Interface: Three Systems
Infection at the Wildlife- livestock-human interface: three systems Thesis submitted in accordance with the requirements of the University of Liverpool for the degree of Doctor in Philosophy by Elsa Sandoval Barron 12/4/2017 Abstract Zoonoses involve interactions between at least three species: the pathogen and two hosts, one of which is human and the other a non-human (vertebrate) animal. More than 60% of human infectious diseases are zoonotic, and many have a wildlife host. Urbanisation and human population growth have increased the demand for food and land resources, which have increased interaction between humans, domestic animals and wildlife and thus the potential for cross-species transmission of infections. Most studies of such systems take place in tropical and developing countries where population change and biodiversity makes the emergence of high profile infections (eg Ebola and SARS) more likely. This study, however, focuses on four well known infections within the UK: bovine tuberculosis, water-borne cryptosporidiosis and giardiasis, and campylobacteriosis. The aim of this study was to investigate, using four infectious diseases of economic and public health importance in the UK as study systems, the role of wildlife in the epidemiology of multihost, zoonotic infections. Bovine tuberculosis (bTB) is an important zoonosis in many parts of the world, but human infection is rare in the UK owing to a policy of ‘test and cull’ in cattle and pasteurisation of milk. However, there has been an epidemic of bTB in British cattle in recent decades, the control of which is complicated by infection in badgers (Meles meles) and controversy over the control of wildlife infection. -
MYCOBACTERIA.Pdf
MYCOBACTERIA Mycobacteria are widespread organisms, typically living in and food sources. Tuberculosis and the leprosy organisms are obligate parasites and are not found as free-living members of the genus. Mycobacteria are aerobic and nonmotile bacteria (except for the species Mycobacterium marinum, which has been shown to be motile within macrophages) that are characteristically acid fast. Mycobacteria have an outer membrane. They do not have capsules, and most do not form endospores. The distinguishing characteristic of all Mycobacterium species is that the cell wall is thicker than in many other bacteria, which is hydrophobic, waxy, and rich in mycolic acids/mycolates. The cell wall consists of the hydrophobic mycolate layer and a peptidoglycan layer held together by a polysaccharide, arabinogalactan. The cell wall makes a substantial contribution to the hardiness of this genus. The biosynthetic pathways of cell wall components are potential targets for new drugs for tuberculosis. Fig. 1 Mycobacterial cell wall: 1-outer lipids, 2-mycolic acid, 3-polysaccharides (arabinogalactan), 4-peptidoglycan, 5-plasma membrane, 6-lipoarabinomannan (LAM), 7- phosphatidylinositol mannoside, 8-cell wall skeleton. NONTUBERCULOUS MYCOBACTERIA – RUNYON CLASSIFICATION Runyon classification is a system of identifying mycobacteria on the basis of pigmentation and growth condition of the organisms. The Runyon classification of nontuberculous mycobacteria based on the rate of growth, production of yellow pigment and whether this pigment was produced in the dark or only after exposure to light. It was introduced by Ernest Runyon in 1959 (Fig. 111). On these bases, the nontuberculous mycobacteria are divided into four groups: Photochromogens (Group I) - produce nonpigmented colonies when grown in the dark and pigmented colonies only after exposure to light and reincubation (1M. -
Cell Structure and Function in the Bacteria and Archaea
4 Chapter Preview and Key Concepts 4.1 1.1 DiversityThe Beginnings among theof Microbiology Bacteria and Archaea 1.1. •The BacteriaThe are discovery classified of microorganismsinto several Cell Structure wasmajor dependent phyla. on observations made with 2. theThe microscope Archaea are currently classified into two 2. •major phyla.The emergence of experimental 4.2 Cellscience Shapes provided and Arrangements a means to test long held and Function beliefs and resolve controversies 3. Many bacterial cells have a rod, spherical, or 3. MicroInquiryspiral shape and1: Experimentation are organized into and a specific Scientificellular c arrangement. Inquiry in the Bacteria 4.31.2 AnMicroorganisms Overview to Bacterialand Disease and Transmission Archaeal 4.Cell • StructureEarly epidemiology studies suggested how diseases could be spread and 4. Bacterial and archaeal cells are organized at be controlled the cellular and molecular levels. 5. • Resistance to a disease can come and Archaea 4.4 External Cell Structures from exposure to and recovery from a mild 5.form Pili allowof (or cells a very to attach similar) to surfacesdisease or other cells. 1.3 The Classical Golden Age of Microbiology 6. Flagella provide motility. Our planet has always been in the “Age of Bacteria,” ever since the first 6. (1854-1914) 7. A glycocalyx protects against desiccation, fossils—bacteria of course—were entombed in rocks more than 3 billion 7. • The germ theory was based on the attaches cells to surfaces, and helps observations that different microorganisms years ago. On any possible, reasonable criterion, bacteria are—and always pathogens evade the immune system. have been—the dominant forms of life on Earth. -
Chemical Probes to Visualize Bacterial Cell Structure and Physiology
molecules Review From Differential Stains to Next Generation Physiology: Chemical Probes to Visualize Bacterial Cell Structure and Physiology Jonathan Hira 1, Md. Jalal Uddin 1 , Marius M. Haugland 2 and Christian S. Lentz 1,* 1 Research Group for Host-Microbe Interactions, Department of Medical Biology and Centre for New Antibacterial Strategies (CANS), UiT—The Arctic University of Norway, 9019 Tromsø, Norway; [email protected] (J.H.); [email protected] (M.J.U.) 2 Department of Chemistry and Centre for New Antibacterial Strategies (CANS), UiT—The Arctic University of Norway, 9019 Tromsø, Norway; [email protected] * Correspondence: [email protected] Academic Editor: Steven Verhelst Received: 30 September 2020; Accepted: 23 October 2020; Published: 26 October 2020 Abstract: Chemical probes have been instrumental in microbiology since its birth as a discipline in the 19th century when chemical dyes were used to visualize structural features of bacterial cells for the first time. In this review article we will illustrate the evolving design of chemical probes in modern chemical biology and their diverse applications in bacterial imaging and phenotypic analysis. We will introduce and discuss a variety of different probe types including fluorogenic substrates and activity-based probes that visualize metabolic and specific enzyme activities, metabolic labeling strategies to visualize structural features of bacterial cells, antibiotic-based probes as well as fluorescent conjugates to probe biomolecular uptake pathways. Keywords: activity-based probe; antibiotic conjugate; bacterial imaging; bacterial uptake; fluorogenic substrate; metabolic labeling; phenotypic heterogeneity 1. Introduction—From 19th Century Microbiology to Modern Day Chemical Biology If chemical biology can be defined as the ‘interrogation of biological systems with chemical approaches’ [1], we must acknowledge some of the first microbiologists as chemical biologists. -
Bacterial Size, Shape and Arrangement & Cell Structure And
Lecture 13, 14 and 15: bacterial size, shape and arrangement & Cell structure and components of bacteria and Functional anatomy and reproduction in bacteria Bacterial size, shape and arrangement Bacteria are prokaryotic, unicellular microorganisms, which lack chlorophyll pigments. The cell structure is simpler than that of other organisms as there is no nucleus or membrane bound organelles.Due to the presence of a rigid cell wall, bacteria maintain a definite shape, though they vary as shape, size and structure. When viewed under light microscope, most bacteria appear in variations of three major shapes: the rod (bacillus), the sphere (coccus) and the spiral type (vibrio). In fact, structure of bacteria has two aspects, arrangement and shape. So far as the arrangement is concerned, it may Paired (diplo), Grape-like clusters (staphylo) or Chains (strepto). In shape they may principally be Rods (bacilli), Spheres (cocci), and Spirals (spirillum). Size of Bacterial Cell The average diameter of spherical bacteria is 0.5- 2.0 µm. For rod-shaped or filamentous bacteria, length is 1-10 µm and diameter is 0.25-1 .0 µm. E. coli , a bacillus of about average size is 1.1 to 1.5 µm wide by 2.0 to 6.0 µm long. Spirochaetes occasionally reach 500 µm in length and the cyanobacterium Accepted wisdom is that bacteria are smaller than eukaryotes. But certain cyanobacteria are quite large; Oscillatoria cells are 7 micrometers diameter. The bacterium, Epulosiscium fishelsoni , can be seen with the naked eye (600 mm long by 80 mm in diameter). One group of bacteria, called the Mycoplasmas, have individuals with size much smaller than these dimensions. -
Transformation
BNL-71843-2003-BC The Ptieumococcus Editor : E. Tuomanen Associate Editors : B. Spratt, T. Mitchell, D. Morrison To be published by ASM Press, Washington. DC Chapter 9 Transformation Sanford A. Lacks* Biology Department Brookhaven National Laboratory Upton, NY 11973 Phone: 631-344-3369 Fax: 631-344-3407 E-mail: [email protected] Introduction Transformation, which alters the genetic makeup of an individual, is a concept that intrigues the human imagination. In Streptococcus pneumoniae such transformation was first demonstrated. Perhaps our fascination with genetics derived from our ancestors observing their own progeny, with its retention and assortment of parental traits, but such interest must have been accelerated after the dawn of agriculture. It was in pea plants that Gregor Mendel in the late 1800s examined inherited traits and found them to be determined by physical elements, or genes, passed from parents to progeny. In our day, the material basis of these genetic determinants was revealed to be DNA by the lowly bacteria, in particular, the pneumococcus. For this species, transformation by free DNA is a sexual process that enables cells to sport new combinations of genes and traits. Genetic transformation of the type found in S. pneumoniae occurs naturally in many species of bacteria (70), but, initially only a few other transformable species were found, namely, Haemophilus influenzae, Neisseria meningitides, Neisseria gonorrheae, and Bacillus subtilis (96). Natural transformation, which requires a set of genes evolved for the purpose, contrasts with artificial transformation, which is accomplished by shocking cells either electrically, as in electroporation, or by ionic and temperature shifts. Although such artificial treatments can introduce very small amounts of DNA into virtually any type of cell, the amounts introduced by natural transformation are a million-fold greater, and S. -
Structure of Bacterial and Archaeal Cells
© Jones & Bartlett Learning, LLC. NOT FOR SALE OR DISTRIBUTION 4 CHAPTER PREVIEW 4.1 There Is Tremendous Diversity Structure of Among the Bacteria and Archaea 4.2 Prokaryotes Can Be Distinguished by Their Cell Shape and Arrangements Bacterial and 4.3 An Overview to Bacterial and Archaeal Cell Structure 4.4 External Cell Structures Interact Archaeal Cells with the Environment Investigating the Microbial World 4: Our planet has always been in the “Age of Bacteria,” ever since the The Role of Pili first fossils—bacteria of course—were entombed in rocks more than TexTbook Case 4: An Outbreak of 3 billion years ago. On any possible, reasonable criterion, bacteria Enterobacter cloacae Associated with a Biofilm are—and always have been—the dominant forms of life on Earth. —Paleontologist Stephen J. Gould (1941–2002) 4.5 Most Bacterial and Archaeal Cells Have a Cell Envelope “Double, double toil and trouble; Fire burn, and cauldron bubble” is 4.6 The Cell Cytoplasm Is Packed the refrain repeated several times by the chanting witches in Shakespeare’s with Internal Structures Macbeth (Act IV, Scene 1). This image of a hot, boiling cauldron actu- MiCroinquiry 4: The Prokaryote/ ally describes the environment in which many bacterial, and especially Eukaryote Model archaeal, species happily grow! For example, some species can be iso- lated from hot springs or the hot, acidic mud pits of volcanic vents ( Figure 4.1 ). When the eminent evolutionary biologist and geologist Stephen J. Gould wrote the opening quote of this chapter, he, as well as most micro- biologists at the time, had no idea that embedded in these “bacteria” was another whole domain of organisms. -
Emerging Frontiers in Microbiome Engineering.Pdf
Trends in Immunology Review Emerging Frontiers in Microbiome Engineering Marı´a Eugenia Inda,1,2,5 Esther Broset,3,4,5 Timothy K. Lu,1,2,* and Cesar de la Fuente-Nunez3,* The gut microbiome has a significant impact on health and disease and can actively contribute to Highlights obesity, diabetes, inflammatory bowel disease, cardiovascular disease, and neurological disor- Themicrobiomeplaysafunda- ders. We do not yet have the necessary tools to fine-tune the microbial communities that consti- mental role in our health and tute the microbiome, though such tools could unlock extensive benefits to human health. Here, disease. we provide an overview of the current state of technological tools that may be used for micro- biome engineering. These tools can enable investigators to define the parameters of a healthy Engineering the microbiome might enable studying the contribution of microbiome and to determine how gut bacteria may contribute to the etiology of a variety of individual microbes and generating diseases. These tools may also allow us to explore the exciting prospect of developing targeted potential therapies against meta- therapies and personalized treatments for microbiome-linked diseases. bolic (e.g., phenylketonuria and chronic kidney disease), inflamma- tory, and immunological diseases, Modulating the Microbiome: An Emerging Paradigm for Understanding and Treating among others. Human Diseases Current methods for probing the The human body is home to at least as many microbial cells as human cells [1]. However, the most microbiome include fecal micro- salient characteristic of the interaction between microbes and the human body is not the number biota transplantation and the use of of cells involved, but their inextricable link with each other. -
Arrayed Crispri and Quantitative Imaging Describe the Morphotypic Landscape of Essential Mycobacterial Genes
RESEARCH ARTICLE Arrayed CRISPRi and quantitative imaging describe the morphotypic landscape of essential mycobacterial genes Timothy J de Wet1,2*, Kristy R Winkler1,2, Musa Mhlanga2,3, Valerie Mizrahi1,2,4, Digby F Warner1,2,4* 1SAMRC/NHLS/UCT Molecular Mycobacteriology Research Unit, Department of Pathology, University of Cape Town, Cape Town, South Africa; 2Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Cape Town, South Africa; 3Department of Integrative Biomedical Sciences, University of Cape Town, Cape Town, South Africa; 4Wellcome Centre for Infectious Diseases Research in Africa, University of Cape Town, Cape Town, South Africa Abstract Mycobacterium tuberculosis possesses a large number of genes of unknown or predicted function, undermining fundamental understanding of pathogenicity and drug susceptibility. To address this challenge, we developed a high-throughput functional genomics approach combining inducible CRISPR-interference and image-based analyses of morphological features and sub-cellular chromosomal localizations in the related non-pathogen, M. smegmatis. Applying automated imaging and analysis to 263 essential gene knockdown mutants in an arrayed library, we derive robust, quantitative descriptions of bacillary morphologies consequent on gene silencing. Leveraging statistical-learning, we demonstrate that functionally related genes cluster by morphotypic similarity and that this information can be used to inform investigations of gene function. Exploiting this observation, we infer the existence of a mycobacterial restriction- modification system, and identify filamentation as a defining mycobacterial response to histidine *For correspondence: [email protected] (TJW); starvation. Our results support the application of large-scale image-based analyses for [email protected] (DFW) mycobacterial functional genomics, simultaneously establishing the utility of this approach for drug mechanism-of-action studies. -
Mycolic Acid (M4537)
Mycolic acid from Mycobacterium tuberculosis (bovine strain) Catalog Number M4537 Storage Temperature –20 °C CAS RN 37281-34-8 Due to the high lipid content of the cell wall, mycobacteria do not stain well with Gram stain Product Description techniques. Heat and a solvent such as phenol are Among different groups of bacteria (e.g., Gram-positive, required for stains to penetrate mycobacteria. Once Gram-negative, spirochetes, mycobacteria, and stained, however, the bacteria retain the stain even mycoplasma), there are various types of cell envelopes when flooded with mineral acids and alcohols. This that represent a departure from the normal simplicity of ability to retain stains after acid washings defines bacterial cell structure compared to animal cells. acid-fast bacteria. Included among the components of the mycobacterium The impermeable cell wall impedes the entry of cell envelope are the cytoplasmic membrane, the cell nutrients causing mycobacteria to grow slowly, but the wall, and the capsule. The cytoplasmic membrane is a low permeability also contributes to the organism’s high phospholipid bilayer unit membrane similar to that resistance to chemical agents and resistance to found in eukaryotic cells. Overlaying the cytoplasmic lysosomal digestion by phagocytes. membrane is a structure consisting of a number of polymers called the cell wall. As for most types of Successful lysis of Mycobacterium tuberculosis by bacteria, highly crosslinked peptidoglycan is a phagocytes causes the release of mycolic acid. The component of the cell wall. Surrounding the cell wall is mycolic acid molecules bind to receptors on an outer layer called the capsule consisting of macrophages causing them to release cytokines such polysaccharide and protein with traces of lipid.1 as tumor necrosis factor-alpha (TNF-a). -
Demonstration of Pneumococcal Capsule Under Immunoelectron Microscopy
Acta Med. Nagasaki 56: 1-4 MS#AMN 07071 Demonstration of pneumococcal capsule under immunoelectron microscopy Akitoyo ICHINOSE,1 Kiwao WATANABE,2 Masachika SENBA,3 Kamruddin AHMED,4 Koya ARIYOSHI,2 Keizo MATSUMOTO5 1 Electron Microscopy Shop, Central Laboratory, Institute of Tropical Medicine, Nagasaki University, Nagasaki 852-8523, Japan 2 Department of Clinical Medicine, Institute of Tropical Medicine, Nagasaki University, Nagasaki 852-8523, Japan 3 Department of Pathology, Institute of Tropical Medicine, Nagasaki University, Nagasaki 852-8523, Japan 4 Institution of Scientific Research, Social and Environmental Medicine, Oita University, Yufu 879-5593, Japan 5 Professor emeritus, Nagasaki University, Nagasaki 852-8523, Japan It is challenging to demonstrate the capsule of Streptococcus pneumoniae (S. pneumoniae) under immunoelectron micros- copy because of the thick mucopeptide cell wall hampering proper fixation. A novel rapid freeze fixation method was estab- lished to observe the capsule of S. pneumoniae. A strain of serotype 3 of S. pneumoniae isolate was analyzed after rapid freezing. An ethanol freezing-substitution fixing method was applied and immunohistochemical staining with osmium tetroxide was tested. The capsule was confirmed using the serotype 3 specific polyclonal antibodies labeled with colloidal gold particles. To the best of our knowledge, this is the first report of S. pneumoniae capsule by immunoelectron microscope. ACTA MEDICA NAGASAKIENSIA 56: 1-4, 2011 Keywords: Pneumococcal capsule, rapid freeze-substitution fixation method, colloidal gold, electron microscopy Introduction ages of pneumococcal capsule by using quick freezing sub- stitution fixation for the first time. Streptococcus pneumoniae (S. pneumoniae) colonizes in human nasopharynx, which may sequentially invades the lungs, the blood stream, and the central nervous system.