BACTERIAL PROFILES

Universal phylogenetic tree

Classification Systems in the Prokaryote

1.
2. 3.

4.
5.

Microscopic morphology Macroscopic morphology colony appearance Bacterial physiology Serological analysis Genetic and molecular analysis

Bacterial Taxonomy Based on Bergeys Manual

Bergeys

Manual of Determinative Bacteriology five volume resource covering all known prokaryotes
Classification based on genetic information phylogenetic Two domains: Archaea and Bacteria Five major subgroups with 25 different phyla

Species and Subspecies

Species a collection of bacterial cells which share an overall similar pattern of traits in contrast to other bacteria whose pattern differs significantly Strain or variety a culture derived from a single parent that differs in structure or metabolism from other cultures of that species (biovars, morphovars) Type a subspecies that can show differences in antigenic makeup (serotype or serovar), susceptibility to bacterial viruses (phage type) and in pathogenicity (pathotype)

Characteristics of Cells
Eukaryotic cells: animals, plants, fungi, and protists

Contain membrane-bound organelles that compartmentalize the cytoplasm and perform specific functions Contain double-membrane bound nucleus with DNA chromosomes No nucleus or other membrane-bound organelles

Prokaryotic cells: bacteria and archaea

Characteristics of Life

Reproduction and heredity genome composed of DNA packed in chromosomes; produce offspring sexually or asexually Growth and development Metabolism chemical and physical life processes Movement and/or irritability respond to internal/external stimuli; self-propulsion of many organisms Cell support, protection, and storage mechanisms cell walls, vacuoles, granules and inclusions Transport of nutrients and waste
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Comparison of Three Cellular Domains

Structure of a bacterial cell

Prokaryotic Profiles

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Prokaryotic Profiles
Structures

that are essential to the functions of all prokaryotic cells are a cell membrane, cytoplasm, ribosomes, and one (or a few) chromosomes

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External Structures
Appendages

Two major groups of appendages:

Motility flagella and axial filaments (periplasmic flagella) Attachment or channels fimbriae and pili

Glycocalyx

surface coating

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Flagella

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Flagella are filamentous protein structures attached to the cell surface that provide the swimming movement for most motile prokaryotes. Prokaryotic flagella are much thinner than eukaryotic flagella, and they lack the typical 9 + 2 arrangement of microtubules. The diameter of a prokaryotic flagellum is about 20 nanometers, wellbelow the resolving power of the light microscope. The flagellar filament is rotated by a motor apparatus in the plasma membrane allowing the cell to swim in fluid environments. Bacterial flagella are powered by proton motive force (chemiosmotic potential) established on the bacterial membrane, rather than ATP hydrolysis which powers eukaryotic flagella. About half of the bacilli and all of the spiral and curved bacteria are motile by means of flagella. Very few cocci are motile

The flagellar apparatus consists of several distinct proteins: a system of rings embedded in the cell envelope (the basal body), a hook-like structure near the cell surface, and the flagellar filament. The innermost rings, the M and S rings, located in the plasma membrane, comprise the motor apparatus. The outermost rings, the P and L rings, located in the periplasm and the outer membrane respectively, function as bushings to support the rod where it is joined to the hook of the filament on the cell surface. As the M ring turns, powered by an influx of protons, the rotary motion is transferred to the filament which turns to propel the bacterium

Flagella

3 parts:

Filament long, thin, helical structure composed of protein Flagellin Hook curved sheath Basal body stack of rings firmly anchored in cell wall

Rotates 360o Number and arrangement of flagella varies:

Monotrichous, lophotrichous, amphitrichous, peritrichous

Functions in motility of cell through environment

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Flagellar Arrangements
1.

2.
3. 4.

Monotrichous single flagellum at one end Lophotrichous small bunches emerging from the same site Amphitrichous flagella at both ends of cell Peritrichous flagella dispersed over surface of cell; slowest

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Electron micrographs of flagellar arrangements

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Flagellar Responses
Guide bacteria in a direction in response to external stimulus:
Chemical stimuli chemotaxis; positive and negative Light stimuli phototaxis

Signal sets flagella into rotary motion clockwise or counterclockwise:

Counterclockwise results in smooth linear direction run Clockwise tumbles
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The operation of flagella

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Chemotaxis in bacteria

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Periplasmic Flagella
Internal

flagella, enclosed in the space between the outer sheath and the cell wall peptidoglycan Produce cellular motility by contracting and imparting twisting or flexing motion

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Periplasmic flagella

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Fimbriae
Fine,

proteinaceous, hairlike bristles emerging from the cell surface Function in adhesion to other cells and surfaces

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Fimbriae

Fimbriae and Pili are interchangeable terms used to designate short, hair-like structures on the surfaces of prokaryotic cells. Like flagella, they are composed of protein. Fimbriae are shorter and stiffer than flagella, and slightly smaller in diameter. Generally, fimbriae have nothing to do with bacterial movement (there are exceptions). Fimbriae are very common in Gram negative bacteria, but occur in some archaea and Gram-positive bacteria as well. Fimbriae are most often involved in adherence of bacteria to surfaces, substrates and other cells in nature. In E.coli, a specialized type of pilus, the F or sex pilus, mediates the transfer of DNA between mating bacteria during the process of conjugation, but the function of the smaller, more numerous common pili is quite different.

 Rigid

tubular structure made of pilin protein Found only in gram-negative cells Function to join bacterial cells for partial DNA transfer called conjugation

Pili

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PILI Common pili (almost always called fimbriae) are usually involved in specific adherence (attachment) of procaryotes to surfaces in nature. In medical situations, they are major determinants of bacterial virulence because they allow pathogens to attach to (colonize) tissues and/or to resist attack by phagocytic white blood cells. For example, pathogenic Neisseria gonorrhoeae adheres specifically to the human cervical or urethral epithelium by means of its fimbriae; enterotoxigenic strains of E. coli adhere to the mucosal epithelium of the intestine by means of specific fimbriae; the M-protein and associated fimbriae of Streptococcus pyogenes help the bacterium resist engulfment by phagocytes.

 Capsules
Most

procaryotes contain some sort of a polysaccharide layer outside of the cell wall polymer called a capsule A true capsule is a discrete detectable layer of polysaccharides deposited outside the cell wall. A less discrete structure or matrix which embeds the cells is a called a slime layer. A type of capsule found in bacteria called a glycocalyx is a thin layer of tangled polysaccharide fibers.

Glycocalyx

Coating of molecules external to the cell wall, made of sugars and/or proteins Two types:
1. Slime layer - loosely organized and attached 2. Capsule - highly organized, tightly attached

Functions:
Protect cells from dehydration and nutrient loss Inhibit killing by white blood cells by phagocytosis, contributing to pathogenicity Attachment - formation of biofilms
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Capsules have several functions and often have multiple functions in a particular organism. Like fimbriae, capsules, slime layers, and glycocalyx often mediate adherence of cells to surfaces. Capsules also protect bacterial cells from engulfment Capsules in certain soil bacteria protect them from perennial effects of drying or desiccation. Capsular materials (e.g. dextrans) may be overproduced when bacteria are fed sugars to become reserves of carbohydrate for subsequent metabolism. Another important characteristic of capsules may be their ability to block some step in the phagocytic process and thereby prevent bacterial cells from being engulfed or destroyed by phagocytes.

CELL WALL
1.They are an essential structure for viability, as described above. 2. They are composed of unique components found nowhere else in nature. 3. They are one of the most important sites for attack byantibiotics. 4. They provide ligands for adherence and receptor sites for drugs or viruses. 5. They cause symptoms of disease in animals. 6. They provide for immunological distinction and immunological variation among strains of bacteria.

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The Cell Wall Most prokaryotes have a rigid cell wall. The cell wall is an essential structure that protects the cell protoplast from mechanical damage and from osmotic rupture or lysis. Prokaryotes usually live in relatively dilute environments such that the accumulation of solutes inside the prokaryotic cell cytoplasm greatly exceeds the total solute concentration in the outside environment. Thus, the osmotic pressure against the inside of the plasma membrane may be the equivalent of 10-25 atm. Since the membrane is a delicate, plastic structure, it must be restrained by an outside wall made of porous, rigid material that has high tensile strength. Such a material is murein, the ubiquitous component of bacterial cell walls.

The cell walls of all Bacteria contain a unique type of peptidoglycan called murein. Peptidoglycan is a polymer of disaccharides (a glycan) cross-linked by short chains of amino acids (peptides), and many types of peptidoglycan exist. All Bacterial peptidoglycans contain N-acetylmuramic acid (NAM) which is the definitive component of murein. The cell walls of Archaea may be composed of protein, polysaccharides, or peptidoglycan-like molecules, but never do they contain murein. In the Gram positive Bacteria (those that retain the purple crystal violet dye when subjected to the Gram-staining procedure) the cell wall is thick (15-80 nanometers), consisting of several layers of peptidoglycan. In the Gram negative Bacteria is composed of a single layer of peptidoglycan surrounded by a membranous structure called the outer membrane. The outer membrane of Gram-negative bacteria invariably contains a unique component, lipopolysaccharide (LPS or endotoxin), which is toxic to animals. Peptidoglycan structure and arrangement in E. coli is representative of all Enterobacteriaceae, and many other Gram-negative bacterial cell wall

 The

glycan backbone is a repeat polymer of two amino sugars, N-acetylglucosamine (NAG) (GlNAc) and N-acetylmuramic acid (MurNAc) (NAM) Attached to the MurNAc /NAM is a tetrapeptide consisting of L-ala-D-glu-DAP-Dala - tetrapeptide side chains may be linked to one another by an interpeptide bond between DAP on one chain and D-ala on the other.

D-ala in an adjacent tetrapeptide side chain. Gram-positive peptidoglycans differ from species to species, mainly in regards to the amino acids in the third position of the tetrapeptide side chain and in the amino acid composition of the interpeptide bridge. Gram-negative bacteria may contain a single monomolecular layer of murein in their cell walls while Gram-positive bacteria are thought to have several layers or wraps of peptidoglycan.
Closely associated with the layers of peptidoglycan in Gram positive bacteria are a group of molecules called teichoic acids. Teichoic acids are linear polymers of polyglycerol or polyribitol substituted with phosphates and a few amino acids and sugars. The teichoic acid polymers are occasionally anchored to the plasma membrane (called lipoteichoic acids) apparently directed outward at right angles to the layers of peptidoglycan. The functions of teichoic acid are not known. They are essential to viability of Gram-positive bacteria in the wild.

 Another

theory is that teichoic acids are in some way involved in the regulation and assembly of muramic acid subunits on the outside of the plasma membrane. There are instances, particularly in the Streptococci, where in teichoic acids have been implicated in the adherence of the bacteria to tissue surfaces

Gram-Negative Cell Wall

Composed of an outer membrane and a thin peptidoglycan layer Outer membrane is similar to cell membrane bilayer structure Outermost layer contains lipopolysaccharides and lipoproteins (LPS)

Lipid portion (endotoxin) may become toxic when released during infections May function as receptors and blocking immune response Contain porin proteins in upper layer regulate molecules entering and leaving cell

Bottom layer is a thin sheet of peptidoglycan

Periplasmic space above and below peptidoglycan
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Gram-Positive Cell Wall

Thick,

homogeneous sheath of peptidoglycan

20-80 nm thick Includes teichoic acid and lipoteichoic acid: function in cell wall maintenance and enlargement during cell division; move cations across the cell envelope; stimulate a specific immune response Some cells have a periplasmic space, between the cell membrane and cell wall

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The Outer Membrane of Gram-negative Bacteria Of special interest as a component of the Gram-negative cell wall is the outer membrane, a discrete bilayered structure on the outside of the peptidoglycan sheet. For the bacterium, the outer membrane is first and foremost a permeability barrier, but primarily due to its lipopolysaccharide content, it possesses many interesting and important characteristics of Gram-negative bacteria. The outer membrane is a lipid bilayer intercalated with proteins, superficially resembling the plasma membrane. The inner face of the outer membrane is composed of phospholipids similar to the phosphoglycerides that compose the plasma membrane. The outer face of the outer membrane may contain some phospholipid, but mainly it is formed by a different type of amphiphilic molecule which is composed of lipopolysaccharide (LPS). Outer membrane proteins usually traverse the membrane and in one case, anchor the outer membrane to the underlying peptidoglycan sheet.

The Lipid A head of the molecule inserts into the interior of the membrane, and the polysaccharide tail of the molecule faces the aqueous environment. Where the tail of the molecule inserts into the head there is an accumulation of negative charges such that a magnesium cation is chelated between adjacent LPS molecules. This provides the lateral stability for the outer membrane, and explains why treatment of Gram negative bacteria with a powerful chelating agent, such as EDTA, causes dispersion of LPS molecules. Bacterial lipopolysaccharides are toxic to animals. When injected in small amounts endotoxins activate macrophages to produce pyrogens, activate the complement cascade causing inflammation, and activate blood factors resulting in intravascular coagulation and hemorrhage. Endotoxins may play a role in infection by any Gram-negative bacterium. The toxic component of endotoxin (LPS) is Lipid A. The O-specific polysaccharide may provide ligands for bacterial attachment and confer some resistance to phagocytosis.

Membrane Lipids

Cell membrane structure

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Cell Membrane Structure

Phospholipid

bilayer with embedded proteins fluid mosaic model Functions in:

Providing site for energy reactions, nutrient processing, and synthesis Passage of nutrients into the cell and the discharge of wastes
Cell membrane is selectively permeable

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Component & Function Lipopolysaccharide (LPS) -Permeability barrier Mg++ bridges - Stabilizes LPS and is essential for its permeability characteristics Braun lipoprotein - Anchors outer membrane to peptidoglycan (murein) Omp C and Omp F - proteins that form pores or porins channels through outer membrane for passage of hydrophilic molecules Omp A protein - provides receptor for some viruses and bacteriocins; stabilizes mating cells during conjugation

Nontypical Cell Walls

Some

bacterial groups lack typical cell wall structure, i.e., Mycobacterium and Nocardia
Gram-positive cell wall structure with lipid mycolic acid (cord factor)
Pathogenicity and high degree of resistance to certain chemicals and dyes Basis for acid-fast stain used for diagnosis of infections caused by these microorganisms

Some

have no cell wall, i.e., Mycoplasma

Cell wall is stabilized by sterols Pleomorphic

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Extreme variation in shape of Mycoplasma pneumoniae

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Bacterial Internal Structures

Cell

cytoplasm:

Dense gelatinous solution of sugars, amino acids, and salts 70-80% water
Serves as solvent for materials used in all cell functions

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Bacterial Internal Structures

Chromosome

Single, circular, double-stranded DNA molecule that contains all the genetic information required by a cell Aggregated in a dense area called the nucleoid
DNA is tightly coiled

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Chromosome structure

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Bacterial Internal Structures

Plasmids

Small circular, double-stranded DNA Free or integrated into the chromosome Duplicated and passed on to offspring Not essential to bacterial growth and metabolism May encode antibiotic resistance, tolerance to toxic metals, enzymes, and toxins Used in genetic engineering - readily manipulated and transferred from cell to cell

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 Ribosomes

Made of 60% ribosomal RNA and 40% protein Consist of two subunits: large and small Prokaryotic differ from eukaryotic ribosomes in size and number of proteins Site of protein synthesis

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Prokaryotic ribosome

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 Inclusions

and granules

Intracellular storage bodies Vary in size, number, and content Bacterial cell can use them when environmental sources are depleted Examples: glycogen, poly bhydroxybutyrate, gas vesicles, sulfur and phosphate granules, particles of iron oxide
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 Cytoskeleton

Many bacteria possess an internal network of protein polymers that is closely associated with the cell wall

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Bacterial Shapes, Arrangements, and Sizes

Vary

in shape, size, and arrangement but typically described by one of three basic shapes:

Coccus spherical Bacillus rod

Coccobacillus very short and plump Vibrio gently curved

Spirillum helical, comma, twisted rod,

Spirochete spring-like

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Common bacterial shapes

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Comparison of Spiral-Shaped Bacteria

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Bacterial Arrangements

Arrangement of cells is dependent on pattern of division and how cells remain attached after division:

Cocci:
Singles Diplococci in pairs Tetrads groups of four Irregular clusters Chains Cubical packets (sarcina)

Bacilli:
Diplobacilli Chains Palisades

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Arrangement of bacilli

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Biofilm on a catheter

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Archaea: The Other Prokaryotes

Constitute third Domain Archaea Seem more closely related to Domain Eukarya than to bacteria Contain unique genetic sequences in their rRNA Have unique membrane lipids and cell wall construction Live in the most extreme habitats in nature, extremophiles Adapted to heat, salt, acid pH, pressure, and atmosphere Includes: methane producers, hyperthermophiles, extreme halophiles, and sulfur reducers
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Archaea

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Comparison of Three Cellular Domains

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