Chair of Microbiology, Virology, and Immunology THE HISTORY OF MICROBIOLOGY. CLASSIFICATION AND STRUCTURE OF MICROORGANISMS. Lecturer As. O. B Kuchmak Lecture schedule 1. 2. 3. 4. History of Microbiology. Classification of bacteria. Structure of bacterial cell The physiology of microorganisms. Growth, reproduction and respiration of bacteria. Why Study Microbiology? Pharmaceuticals Vaccines/Antibiotics Biotechnology Bioremediation Pathogenicity Fundamental Biology Microbiology The study of microorganisms Microorganisms living things too small to be seen with the unaided eye Microorganisms = Microbes Microorganisms Divided into six groups Bacteria Archae Algae Fungi Protozoa Viruses History of Microbiology Microbes discovered >300yrs Known to man during the mid 1800s Period of progress began & continues to the present Periods of microbiology development Morphologic Physiologic Prophylactic Morphological period in microbiology history (XVII middle of age). It is also called micrographycal period, as the study of microorganism came only to description of their dimensions and forms. Biological properties and their significances for man still a long time remained incomprehensible. Experimental phase Leeuwenhoek Pasteur Koch Lister Before 17th century, study of microbiology was hampered by the lack of appropriate tools to observe microbes. Robert Hooke: In 1665 built a compound light microscope and used it to observe thin slices of cork. Coined the word cell. Anton van Leeuwenhoeck: In 1673 was the first person to observe live microorganisms which he called “animalcules” (bacteria, protozoa), using single-lens microscopes that he designed. He observed 50,000 different specimens, reported findings to the Royal Society of London Jenner ( 1796) Smallpox immunity / Vaccine Edward Pasteur’s Contributions: Pasteurization: Developed a process in which liquids are heated (at 65oC) to kill most bacteria responsible for spoilage. Disease Causes: Identified three different microbes that caused silkworm diseases. Vaccine: Developed a vaccine for rabies from dried spinal cords of infected rabbits. Directed Pasteur Institute until his death in 1895. French Chemist Pasteur (1861) Joseph Lister (1859): Used disinfectant to treat surgical wounds, greatly reducing infection rates. Considered the father of antiseptic surgery. Robert Koch (1876): First person proved that microorganisms caused diseases Only specific microorganisms caused specific diseases Studied anthrax affects cattle & humans Proved that Bacillus anthracis causes anthrax in cattle. Later identified bacterium that causes tuberculosis. Study of viruses Iwanoski (1892) studying diseases of tobacco plants Discovered TMV Filterable viruses Prophylactic period After 1914 Classic Metchnikov's researches defined a prophylactic period in microbiology history. The Patriarch of world and Ukrainian microbiology - I. Metchnikov Paul Ehrlich (1910): Search for “magic bullet”. Discovered salvarsan, an arsenic derivative, was effective against syphilis. (1st synthetic drug to come in to widespread use) Alexander Fleming (1928): Discovered that penicillin produced by the mold Penicillium notatum was able to prevent microbial growth. Penicillin came into use 10 yrs later -By the 1940s known as the “wonder drug” Rene Dubos (1939): Discovered two antibiotics (gramidin and tyrocidine) produced by bacterium (Bacillus brevis). Classifications system. Bergey's Manual of Determinative Bacteriology – the "bible" of bacterial taxonomy. There are such levels of microorganisms’ organization: Species – Genus – Family – Class – Division – Kingdom 35 of the major groups of bacteria are distinguished primarily on morphological characteristics, namely: cell shapes (rods, cocci, curved, or filament forming); spore production; staining reactions; motility. Other groups are defined based on their metabolism, or combinations of morphological and physiological characteristics. Some of the Major Groups of Bacteria in Bergey's Manual Very slender rods that are helically coiled around a central axial filament; Spirochetes includes the bacteria that cause syphilis and Lyme disease Bacteria that have a cell wall structure Gram-positive that results in their staining bluepurple by the Gram stain procedure cocci and that are spherical; include the streptococci and staphylococci Bacteria that form heat-resistant bodies Endosporeforming rods and called endospores within their cells; include the bacteria that cause gas cocci gangrene, botulism, tetanus, and anthrax Bacteria (Sing. Bacterium) Small, single-celled (unicellular) organisms. Procaryotes: “Before nucleus”. Lack the following structures: Nuclear membrane around DNA Membrane bound organelles Mitochondria Chloroplasts Golgi apparatus Endoplasmic Lysosomes reticulum Property Prokaryotic Eukaryotic 0.5 - 10μm 5-100μm Present in all bacteria Except in Mycoplasma Present in Fungi & Algae No Sterol Except in Mycoplasma Has sterols 4- Nuclear Membrane Absent Present 5- Nucleus Absent Present 6- Chromosome Single chromosom [not associtred with protein More than one [associated with histone] 7- Mitochondria Absent Present Sedimentation coefficient 70S Sedimentation coefficient 80S Asexual (binary fusion) Sexual & Asexual Bacteria, Chlamydia, Rickettsiae Fungi & Protozoa 1- Size 2- Cell Wall 3- Cytoplasmic Membrane 8- Ribosome 9- Reproduction 10- Example Bacterial Identification and Classification Shape - cocci, bacilli, spiral Arrangement - single, pairs, chains, clusters Size Gram-positive vs. Gram-negative Aerobic vs. anaerobic Physical/structural characteristics Biochemical characteristics DNA analysis The Dimension of Bacteria Relative size of a bacterial cell compared to other cells including viruses. Gross morphology of bacteria Size - 1 to 10 mm - aids in identification Shape Coccus - round Bacillus - rod - Fusiform, coccobacilli Spirillum - corkscrew - Flexible, undulating - spirochetes - Curved - vibrios Cocci groupings Coccus Diplococcus Streptococcus Tetrad Sarcinae Staphylococcus Chains of cocci Streptococcus pyogenes Clusters of cocci Staphylococcus aureus Sputum smear Bacillus shaped bacteria Pseudomonas aeruginosa Escherichia coli Bacillus shaped bacteria Fusobacterium Bacillus chains Bacillus anthracis Palisades arrangement Corynebacterium diphtheriae Curved bacteria Vibrio cholerae Campylobacter Curved bacteria Spirilla Borrelia burgdorferi Bacterial surface structures Cell Envelope Cytoplasmic Cell membrane wall Cell wall-less bacteria No peptidoglycan layer Cell membrane contains sterols for stability Mycoplasma pneumoniae Cytoplasmic Membrane Phospholipid bilayer “Fluid mosaic” model Embedded proteins for active transport Enzymes for energy generation Photosynthetic pigments Cell membrane Peripheral Membrane Protein Phospholipid Integral Membrane Protein Peripheral Membrane Protein Function of Cytoplasmic Membrane Selective permeability to different molecules. Active transport aided by permease. Play a role in DNA replication. Cell wall biosynthesis. Mesosomes ----- cell division. Cell wall Two major groups of bacteria based on structure of cell wall Gram positive Thick peptidoglycan layer Gram negative Thin peptidoglycan layer Outer membrane containing LPS Gram stain is crucial first step toward identification Peptidoglycan (cell wall) Cell Wall Gram positive cell wall Thick peptidoglycan (PG) layer Acidic polysaccharides Teichoic acid and lipoteichoic acid Gram-negative cell wall Thin peptidoglycan (PG) layer Lipopolysaccharide layer Porins Periplasmic space Gram-positive cell envelope Gram-negative cell envelope Cell Wall Structures Structures associated with gram-positive and gram-negative cell walls. Function of Cell Wall Maintenance of the shape (due to rigidity of peptidoglycan). Protects the cytoplasmic membrane cell contents Rigidity Cell wall is osmotically insensitive Hypotonic solution – cell burst. Hypertonic solution – cell shrank. Isotonic solution – bacteria is life. L Forms Mutations can cause some bacteria to lose the ability to synthesize the cell wall and are called L forms. Capsules are important for Adhesion (Associated with virulence in bacteria) Avoidance of immune response (Protects bacteria from phagocytic cells) Protection from dehydration Capsule Streptococcus pneumoniae Klebsiella pneumoniae Bacillus anthracis External structures Pili (Fimbriae) play roles in Adhesion Exchange of genetic material Avoidance of immune response Flagella are important for Motility (dispersal) Antigenic determinant (“H” antigens) Number and location species specific Pili and flagella Fimbriae are smaller than flagella and are important for attachment. Salmonella Pili Pili enable conjugation to occur, which is the transfer of DNA from one bacterial cell to another (“mating”). Flagellar Structure Three components of a flagellum: filament, hook and basal body It composed of protein subunits called flagellin. Flagellar Arrangement (a) Monotrichous (c) Amphitrichous (b) Lophotrichous (d) Peritrichous Bacterial Motility The rotation of the flagella enables bacteria to be motile. Internal Structures Cytoplasm Genome Inclusion bodies Endospore Cytoplasm Gelatinous solution containing water, nutrients, proteins, and genetic material Site for cell metabolism Chemical Analysis of Microbial Cytoplasm 70% water Proteins 96% of cell is composed of 6 elements: carbon hydrogen oxygen phosphorous sulfur nitrogen 69 Bacterial Genome Most bacteria contain a single circular double strand of DNA called a nucleoid. Prokaryotic Ribosome A ribosome is a combination of RNA and protein, and is the site for protein synthesis Composed of large (50S) and small (30S) subunits S = Svedverg unit, measures molecular size Inclusion Bodies Inclusion bodies enable a cell to store nutrients and to survive in nutrient depleted environments Bacterial Spores Some bacteria, notably those of the genera Bacillus and Clostridium, develop a highly resistant resting phase or endospore that does not grow or reproduce and exhibit absolute dormancy (not detectable metabolism). Endospores Bacillus anthracis Vegatitive form The bacteria actively growing, non spore stage of a bacterium. Sporulation: Formed on exposure to unfavorable condition,E.g., Nutrient depletion Changes Moisture, Temperature, pH or Oxygen tension Spore requires 10-15 hours to form. Endospore formation Germination Mature endoscope are metabolically inert Changes in the environment Retuning to vegetative state within 15 minutes. In the process of germination the spores absorb water and swell, the protective coat disintegrates and a single vegatitive cell emerges.