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Morphology and structure of bacteria Prof. Marianna Murdjeva, MD, PhD Department of Microbiology and Immunology Medical University-Plovdiv Lecture course in microbiology Summer term Bacterial Morphology studies: • • • • Bacterial Shape Bacterial Size Bacterial Cell Arrangement Gram staining According to their shape bacteria are: • Round (cocci) – staphylococci, streptococci – diplococci (pneumococci, meningococci) • Rod-shaped (rods) – non-spore forming (M. tuberculosis, C. diphtheriae) – spore-forming (bacilli) - clostridia, B. anthracis • Spiral (spirilli) – vibrios (V. cholerae) – 1 curve – spirilli (Helicobacter) – 2 curves – spirochaetes: Treponema, Leprospira, Borrelia – many curves According to cell arrangement bacteria are: • Single – monococci • Pairs – diplococci (pneumococci, meningococci) – diplobacteria (Klebsiella) • Tetrads (sarcina) • Chains (streptococci, B. anthracis) • Clusters (staphylococci) Bacteria are measured in micrometers (m) • • • Small (0.2-0.3 m). Haemophillus, Brucella Medium (0.5-2 m). Staphylococci, Streptococci, E. coli Large (3-10 m). Clostridia, B. anthracis Bacterial structure: • Cell envelope = cell wall (CW)+cytoplasmic membrane (CM) • Cytoplasmic components: - core material (nucleoid) –N - ribosomes (Ri) - inclusions • External structures - capsule - flagella and pilli - spores • Essential (obligatory) organels - CW, CM, N, Ri • Non-essential (additional) organels – capsule, flagella, pilli, spores Bacterial Cell wall contains Peptidoglycan: 1. glycan part: N-acetyl glucosamine N-acetyl muramic acid -1,4 glycoside link 2. peptide part Difference b/n Gram+ and Gram- cell wall Gram – Cell Wall Gram+ Cell Wall • • Thick layer of peptidoglycan Negatively charged teichoic acid on surface • • Thin peptidoglycan Outer membrane: - О Ag, LPS and core oligosaccharide. – LPS – Ag and toxic properties. Lipid А (endotoxin). Causes Endotoxic shock. – Porins. • Periplasmic space Function of CW in B • Some bacterial groups lack typical cell wall structure i.e. Mycobacterium and Nocardia: – They have Gram-positive cell wall structure with lipid mycolic acid (cord factor) which are responsible for pathogenicity and high degree of resistance to certain chemicals and dyes • Some have no cell wall i.e. Mycoplasma: – their cell structure is stabilized by sterols – they are pleomorphic • Lysozyme digests disaccharide in peptidoglycan. • Penicillin inhibits peptide bridges in peptidoglycan. Bacterial Cytoplasmic membrane (CM) : • • • • • • • Three-layered Target for lipid-dissolving agents and some antibiotics (Polymyxin and Nistatin) Lack of inner membranes Function of bacterial CM : Respiratory (mitochondrial) equivalent Selective permeability Participates in peptidoglycan synthesis and formation of penicillin-binding proteins, necessary for linkage with some antibiotics Participates in chromosome replication and large plasmids Bacterial mesosomes: • Organels, formed by CM folding • Functions: – respiratory (mitochondrial) equivalent – coordination of core material division and cytoplasm during binary fission of bacteria Bacterial cytoplasmic components • Ribosomes – difference with Eu • Inclusions: – volutine (diphtherial bacteria) – glycogen (enteric bacteria) – others • Core (nucleoid)- a single DNA molecule • Extra-chromosomal genetic elements (plasmids, bacteriophages) Bacteria may have non-essential structures: • • • • Capsule Flagella Pilli Spores Bacterial capsule: • • • • May be real (S. рneumoniae, B. anthracis), slime (S. mutans), or microcapsule (S. typhi) Structure – polysaccharide (S. pneumoniae) – polypeptide (B. anthracis) Staining (Klett, Neuffeld) Function: – protection (virulence factor) – Ag properties (K Ag) Flagella: • • • Structure – 3 parts: - filament – long, thin, helical structure composed of protein flagellin - hook- curved sheath - basal body – stack of rings firmly anchored in cell wall Composition – protein (flagellin) Function – motion, virulence factor, antigenic and receptor Pilli (fimbriae) • • Types – common (adhesins) – sex (participate in conjugation) Composition - protein (pillin=fimbrillin) Function: – adhesion to cells (gonococci, E. coli) – participation in conjugative transfer – Ag properties (F Ag) Bacterial spores • Formation – at high temperature or dehydration • Structure and composition – less water, thicker wall • Types: – according to location in the cell: central (B. anthracis), terminal(С. tetani), subterminal (C. perfringens) – according to shape: round or oval – according to their capacity to deform the cell: deforming (Clostridium) and non-deforming (B. anthracis) Sporulation Microscopy: The Instruments Brightfield Microscopy Darkfield Microscopy • • • Simplest of all the optical microscopy illumination. techniques Dark objects are visible against a bright background. Light objects visible against dark background. used to enhance the contrast in unstained samples. Instrument of choice for spirochetes • • Microscopic staining methods for bacterial structure: – simple (Loffler, Pfeiffer) – differential (Gram, Neisser, Zhiel-Neelsen, Moller, Klett) Fluorescence Microscopy Electron Microscopy: for Detailed Images of Cell Parts • Uses UV light. • Uses electrons, electromagnetic lenses, and fluorescent screens • Fluorescent substances absorb UV light and emit visible light. • • Cells may be stained with fluorescent chemicals (fluorochromes). Electron wavelength ~ 100,000 x smaller than visible light wavelength Specimens may be stained with heavy metal salts •