Bacterial Cell Structure Dr. Majid Zare Bidaki (PhD in Microbiology, Assistant professor) Birjand University of Medical Science References 1. Jawetz, Melnick, & Adelberg; Medical Microbiology, 24th ed. 2. Zinsser, Hans, And Joklik, Wolfgang K.; Medical Microbiology, 17th ed. 3. Baron, Samuel; Medical Microbiology, 4th ed. Antonie van Leeuwenhoek First to observe living microbes his single-lens magnified up to 300X (1632-1723) 3 4 Louis Pasteur (1822-1895) Showed microbes caused fermentation & spoilage Disproved spontaneous generation of microbes Developed aseptic techniques. Developed a rabies vaccine. 5 Robert Koch Proposed: Germ theory of disease Developed: pure culture methods. Identified: cause of anthrax, TB, & Cholera. (1843-1910) 6 Acellular and cellular Microorganisms Acellular: Viruses Viroids Prions Cellular: Bacteria fungi Protista: Protozoa & algae helminths (worms) Prokaryotic or Eukaryotic 8 Prokaryotes vs Eukaryotes Size smaller Larger Nucleus - + Organelles - + Chromosomes 1 circular Multiple, linear Ribosomes Larger 80s smaller 70s r In prokaryotes against Eukaryotes, cell membranes lack sterols (e.g. cholesterol) Scientific nomenclature Binomial (scientific) nomenclature Gives each microbe 2 names Genus - noun, always capitalized species - adjective, lowercase Both italicized or underlined – Staphylococcus aureus (S. aureus) – Escherichia coli (E. coli) 10 Bacterial shapes Cytoplasmic membrane Protoplast Spheroplast L forms Gram positive Gram negative Lipopolysaccharide O-antigen Highly variable Core • Heptoses • Ketodeoxyoctonic acid Lipid A • Glucosamine disaccharide • Beta hydroxy fatty acids (Hydroxy myritic Acid) LPS function Endotoxins Exotoxins Peptidoglycan Gram positive Gram negative 4 groups based on cell wall composition 1. Gram positive cells 2. Gram negative cells 3. Bacteria without cell walls 4. Bacteria with chemically unique cell walls Gram positive wall Gram negative cell wall Lipoteichoic acid Peptidoglycan-teichoic acid Cytoplasmic membrane Cytoplasm Porin Outer Membrane lipoprotein Inner (cytoplasmic) membrane Cytoplasm Lipopolysaccharide Gram Positive Cell Envelope Peptidoglycan-teichoic acid Lipoteichoic acid r r r Cytoplasmic membrane r r r r r r r Bacteria classification based on cell wall structure Grasilicutes (Gram Negative) Firmicutes (Gram Positive) Tenricutes (with no Cell wall) Mendosicutes (with no Peptidoglycan in cell wall) Major Taxonomic Groups of Bacteria Gracilicutes – gram-negative cell walls, thinskinned Firmicutes – gram-positive cell walls, thick skinned Tenericutes – lack a cell wall & are soft Mendosicutes – archaea, primitive procaryotes with unusual cell walls & nutritional habits Capsule 2 types 1. Macro capsule - highly organized, tightly attached 2. Micro capsule, Slime layer or Glycocalyx loosely organized and attached Functions attachment inhibits killing by white blood cells Receptor (K antigen) 2 Types of Capsule Biofilms Flagella Fimbrea (Pili) Adhesion to other cells and surfaces Structure Pili & Sex pili Rigid tubular structure made of pielin protein Found mostly in Gram negative cells Functions Adhesion joins bacterial cells for DNA transfer (Conjugation) Conjugation Cytoplasm dense gelatinous solution of sugars, amino acids, & salts 70-80% water serves as solvent for materials used in all cell functions Chromosome single, circular, double-stranded DNA molecule that contains all the genetic information required by a cell DNA is tightly coiled around a protein, aggregated in a dense area called the nucleoid plasmids small circular, double-stranded DNA free or integrated into the chromosome duplicated and passed on to offspring not essential to bacterial growth & metabolism may encode antibiotic resistance, tolerance to toxic metals, enzymes & toxins used in genetic engineering- readily manipulated & transferred from cell to cell Ribosomes made of 60% ribosomal RNA & 40% protein consist of 2 subunits: large (50 S) & small (30 S) procaryotic differ from eucaryotic ribosomes in size & number of proteins site of protein synthesis All bacterial cells have ribosomes. Inclusions, granules intracellular storage bodies vary in size, number & content bacterial cell can use them when environmental sources are depleted Examples: glycogen, sulfur and polyphosphate granules, poly-b-hydroxybutyrate, gas vesicles for floating. endospores Important components in endospore: Calcium Dipicolinic Acid The Endospore structure Spore structure Spherical or Oval Terminal, subterminal or central Bulging or nobulging Growth in Bacteria Temperature Nutrients pH Osmotic pressure Temperature Minimum temperature – lowest temperature that permits a microbe’s growth and metabolism Maximum temperature – highest temperature that permits a microbe’s growth and metabolism Optimum temperature – promotes the fastest rate of growth and metabolism 3 temperature adaptation groups Bacterial Metabolism Phototroph Photoautotroph (Photolitotroph) Photoheterotroph (Photoorganotroph) Chemotroph Chemoautotroph (Chemolitotroph Chemoheterotroph (Chemoorganotroph) Stages of metabolism in chemoheterotrophic bacteria Digestion Absorption (Passive and active transportation) Preparation for oxidation Oxidation Oxidation & Reduction X e- & H+ Cytochromes, …. Y Oxygen requirements Bacterial growth Binary division G2 G1 G2 G0 G2 G1 G2 Microbial growth calculation b = a X 2n G (Generation time) = T / n (n = The number of generations, T = The total time of growth for the population) The curve of bacterial growth in a closed culture Growth Curve Bacterial growth in a continues culture Continuous Culture, Chemostat Chemostats are a means of keeping a culture in log phase indefinitely. Measuring the bacterial growth • Measuring the mass of bacteria • Measuring the number of bacteria Fermentation Incomplete oxidation of glucose or other carbohydrates in the absence of oxygen Uses organic compounds as terminal electron acceptors Yields a small amount of ATP Production of ethyl alcohol by yeasts acting on glucose Formation of acid, gas & other products by the action of various bacteria on pyruvic acid Fermentation Methods in bacterial identification 1. 2. 3. 4. 5. 6. Microscopic morphology Macroscopic morphology – colony appearance Physiological / biochemical characteristics Chemical analysis Serological analysis Genetic & molecular analysis G + C base composition DNA analysis using genetic probes Nucleic acid sequencing & rRNA analysis Bacterial Colonies • Standard Bacterial Count • Colony-Forming Units • Plaque-Forming Units •Spread Plate • Pour Plate • Soft-Agar Overlay Medium Definition Types based on solidity: 1. Liquid medium (Name broth) BHI, TSB, SF, NB, … 2. Solid medium (Name agar) Blood agar, Nutrient agar, chocolate agar, Columbia agar, EMB 3. Semi-solid medium SIM Culture media General medium Special medium Differential medium Enrichment medium (….. & cold enrichment) Transport medium (Stwart, Carry Blair, …) Galleries Types of culture methods Isolation culture Spread culture Pour plate culture Colony count culture