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CLASSIFICATION AND STRUCTURE OF MICROORGANISMS.
Chair of Microbiology, Virology, and Immunology
Comparison of the three domainsCharacteristic Eubacteria Archaea Eucarya
Cell type Prokaryote Prokaryote Eukaryote
Cell wall Peptidoglycan Varies Varies
Membrane lipids
Unbranched Branched Unbranched
Sensitive to antibiotics?
Yes No No
Circular chromosome?
Yes Yes No (except in mitochondria and chloroplasts)
Histones? No Yes Yes
Prokaryotes
Classification Systems in the Procaryotae
1. Microscopic morphology
2. Macroscopic morphology – colony appearance
3. Physiological / biochemical characteristics
4. Chemical analysis
5. Serological analysis
6. Genetic and molecular analysis• G + C base composition• DNA analysis using genetic probes• Nucleic acid sequencing and rRNA analysis
Bacterial Taxonomy Based on Bergey’s Manual
• Bergey’s Manual of Determinative Bacteriology – five volume resource covering all known procaryotes– classification based on genetic information –
phylogenetic– two domains: Archaea and Bacteria– five major subgroups with 25 different phyla
Taxonomy
• Domain• Kingdom• Phylum• Class• Order• Family• Genus• species
Major Taxonomic Groups of Bacteria
• Vol 1A: Domain Archaea– primitive, adapted to extreme habitats and
modes of nutrition
• Vol 1B: Domain Bacteria • Vol 2-5:
– Phylum Proteobacteria – Gram-negative cell walls
– Phylum Firmicutes – mainly Gram-positive with low G + C content
– Phylum Actinobacteria – Gram-positive with high G + C content
Microbial Phylogeny
• Phylogeny of domain Bacteria– The 2nd edition of Bergey’s Manual of
Systematic Bacteriology divides domain Bacteria into 23 phyla.
Microbial Phylogeny
• Phylogeny of domain Bacteria (cont.) • Phylum Proteobacteria
– The largest group of gram-negative bacteria– Extremely complex group, with over 400 genera and
1300 named species– All major nutritional types are represented: phototrophy,
heterotrophy, and several types of chemolithotrophy– Sometimes called the “purple bacteria,” although very
few are purple; the term refers to a hypothetical purple photosynthetic bacterium from which the group is believed to have evolved
Microbial Phylogeny
• Phylogeny of domain Bacteria (cont.) • Phylum Proteobacteria (cont.)
– Divided into 5 classes: Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Deltaproteobacteria, Epsilonproteobacteria
Microbial Phylogeny
• Phylogeny of domain Bacteria (cont.) • Phylum Proteobacteria (cont.)
– Significant groups and genera include:» The family Enterobacteriaceae, the “gram-negative
enteric bacteria,” which includes genera Escherichia, Proteus, Enterobacter, Klebsiella, Salmonella, Shigella, Serratia, and others
» The family Pseudomonadaceae, which includes genus Pseudomonas and related genera
» Other medically important Proteobacteria include genera Haemophilus, Vibrio, Camphylobacter, Helicobacter, Rickessia, Brucella
Microbial Phylogeny• Phylogeny of domain Bacteria (cont.)
• Phylum Firmicutes– “Low G + C gram-positive” bacteria
– Divided into 3 classes» Class I – Clostridia; includes genera Clostridium and
Desulfotomaculatum, and others
» Class II – Mollicutes; bacteria in this class cannot make peptidoglycan and lack cell walls; includes genera Mycoplasma, Ureaplasma, and others
» Class III – Bacilli; includes genera Bacillus, Lactobacillus, Streptococcus, Lactococcus, Geobacillus, Enterococcus, Listeria, Staphylococcus, and others
Microbial Phylogeny
• Phylogeny of domain Bacteria (cont.) • Phylum Actinobacteria
– “High G + C gram-positive” bacteria– Includes genera Actinomyces, Streptomyces,
Corynebacterium, Micrococcus, Mycobacterium, Propionibacterium
• Phylum Chlamidiae– Small phylum containing the genus Chlamydia
Microbial Phylogeny
• Phylogeny of domain Bacteria (cont.) • Phylum Spirochaetes
– The spirochaetes– Characterized by flexible, helical cells with a modified
outer membrane (the outer sheath) and modified flagella (axial filaments) located within the outer sheath
– Important pathogenic genera include Treponema, Borrelia, and Leptospira
• Phylum Bacteroidetes– Includes genera Bacteroides, Flavobacterium,
Flexibacter, and Cytophyga; Flexibacter and Cytophyga are motile by means of “gliding motility”
Procaryotae Kingdom has 4 Divisions according to the structure of cell wall and Gram staining:
Gracilicutes (gracilis - thin, cutis - skin) – Gram-negative bacteria,
Firmicutes (firmus - firm) – Gram-positive bacteria,
Tenericutes (tener – soft, tender) – microbes without cell wall,
Mendosicutes (mendosus - mistaket) – microbes with atipical peptidoglican
Bacterial Nomenclature
• Binomial naming system– Two word naming system
• First word is genus name– Always capitalized
• Escherichia• Second word is species name
– Not capitalized• coli
• When writing full name genus usually abbreviated– E. coli
• Full name always italicized– Or underlined
Species is population of microbes, which have the only source of origin, common genotype, and during the present stage of evolution are characterized by similar morphological, biochemical, physiological and other signs
If deviations from the typical species properties are found on examination of the isolated bacteria, then culture is considered a subspecies.
Infrasubspecies subdivisions
serovar (antigenic properties)
morphovar (morphological properties)
chemovar (chemical properties)
biovar (biochemical or physiological properties)
pathovar (pathogenic properties)
phagovar (relation to phages)
The term clone was applied to population of cells derived from a single cell
Population is an elementary evolutional unit (structure) of a definite species
The term strain designates a microbial culture obtained from the different sources or from one source but in different time
Or: A subgroup within a species with one or more haracteristics that distinguish it from other subgroups in the species
Morphological Classification of Bacteria
Bacteria (Gk. bakterion - small staff) are unicellular organisms lacking chlorophyll.
Morphologically, bacteria possess four main forms:
spherical (cocci)
rod-shaped (bacteria, bacilli, and clostridia)
spiral-shaped (vibrios, spirilla and spirochaetes)
thread-shaped (non-pathogenic)
Cocci groupings
Coccus
Diplococcus
Streptococcus
Tetrad
Sarcinae
Staphylococcus
Cocci (Gk. kokkos berry). These forms of bacteria are spherical, ellipsoidal, bean-shaped, and lanceolate. Cocci are subdivided into six groups according to cell arrangement, cell division and biological properties
Micrococci (Micrococcus). The cells are arranged singly or irregularly. They are saprophytes, and live in water and in air ( M. roseus, M. luteus, etc.).
Diplococci (Gk. diplos double) divide in one plane and remain attached in pairs. These include: Meningococcus (causative agent of epidemic cerebrospinal meningitis, and gonococcus, causative agent of gonorrhoea and blennorrhoea) Pneumococcus (causative agents of pneumonia)
Streptococci (Gk. streptos curved, kokkos berry) divide in one plane and are arranged in chains of different length. Some streptococci are pathogenic for humans and are responsible for various diseases.
Tetracocci (Gk. tetra four) divide in two planes at right angles to one another and form groups of fours. They very rarely produce diseases in humans.
Staphylococci (Gk. staphyle cluster of grapes) divide in several planes resulting in irregular bunches of cells, sometimes resembling clusters of grapes. Some species of Staphylococci cause diseases in man and animals
Sarcinae (L. sarcio to tie) divide in three planes at right angles to one another and resemble packets of 8, 16 or more cells. They are frequently found in the air. Virulent species have not been encountered
Rods. Rod-shaped forms are subdivided into:
bacteria,
bacilli,
clostridia
Bacteria include those microorganisms which, as a rule, do not produce spores (colibacillus, and organisms responsible for enteric fever, paratyphoids, dysentery, diphtheria, tuberculosis, etc.).
Bacilli and clostridia include organisms the majority of which produce spores (hay bacillus, bacilli responsible for anthrax, tetanus, anaerobic infections, etc.)
According to their arrangement, cylindrical forms can be subdivided into three groups: monobacteria monobacilli
E. coli Y. pestis
C. tetani
C. botulinum
diplobacteria diplobacilli
K. pneumoniae
streptobacteria streptobacilli
Haemophilus ducreyi
(chancroid)
Bacillus anthracis
(anthrax)
Spiral-shaped bacteria
Vibriones (L. vibrio to vibrate) are cells which resemble a comma in appearance. Typical representatives of this group are Vibrio cholerae, the causative agent of cholera, and aquatic vibriones which are widely distributed in fresh water reservoirs.
Spirilla (L. spira coil) are coiled forms of bacteria exhibiting twists with one or more turns. Only one pathogenic species is known {Spirillum minus} which is responsible for a disease in humans transmitted through the bite of rats and other rodents (rat-bite fever, sodoku)
Spirochaetes (L. spira curve, Gk. chaite cock, mane) differ from bacteria in structure with a corkscrew spiral shape
Borrelia. Their cells have large, obtuse-angled, irregular spirals, the number of which varies from 3 to 10. Pathogenic for man are the causative agents of relapsing fever transmitted by lice (Borrelia hispanica), and by ticks (Borrelia persica, etc.). These stain blue-violet with the Romanowsky-Giemsa stain
Leptospira (Gk. leplos thin, speira coil) are characterized by very thin cell structure. The leptospirae form 12 to 18 coils wound close to each other, shaping small primary spirals. The organisms have two paired axial filaments attached at opposite ends (basal bodies) of the cell and directed toward each other.
Leptospira interrogans which is pathogenic for animals and man cause leptospirosis
Treponema (Gk. trepein turn, nema thread) exhibits thin, flexible cells with 6-14 twists. The micro-organisms do not appear to have a visible axial filament or an axial crest when viewed under the microscope
A typical representative is the causative agent of syphilis Treponema pallidum
Properties of prokaryotes and eukaryotes
Prokaryotes Eukaryotes
The nucleoid has no membrane separating it from the cytoplasm
Karyoplasm is separated from the cytoplasm by membrane
Chromosome is a one ball of double twisted DNA threads. Mitosis is absent
Chromosome is more than one, There is a mitosis
DNA of cytoplasm are represented in plasmids
DNA of cytoplasm are represented in organelles
There aren’t cytoplasmic organelle which is surrounded by membrane
There are cytoplasmic organelle which is surrounded by membrane
The respiratory system is localized in cytoplasmic membrane
The respiratory system is localized mitochondrion
There are ribosome 70S in cytoplasm
There are ribosome 80S in cytoplasm
Peptidoglycan are included in cell’s wall (Murein)
Peptidoglycan aren’t included in cell’s wall
The structure of procaryotes
Nucleus. The prokaryotic nucleus can be seen with the light microscope in stained material. It is Feulgen-positive, indicating the presence of DNA. Histonelike proteins have recently been discovered in bacteria and presumably play a role similar to that of histones in eukaryotic chromatin
The DNA is seen to be a single, continuous, "giant" circular molecule with a molecular weight of approximately 3 X 109. The unfolded nuclear DNA would be about 1-3 mm long (compared with an average length of 1 to 2 µm for bacterial cells)
Plasmids: R, Col, Hly, Ent, Sal
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 There may be several different plasmids in one cell and the numbers of each may vary from only one to 100s in a cell
Prokaryotic RibosomeProkaryotic Ribosome
A ribosome (70 S) 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
The 80S ribosomes of eukaryotes are made up of 40S and 60S subunits.
• Storage granules– Metachromatic
granules – Polysaccharide
granules– Lipid inclusions– Sulfur granules– Carboxyzomes – Magnetosomes
• Gas vesicles
Inclusions, granules
Volutin granules
Corynebacterium diphtheriae
Loeffler's technique Neisser's staining
Composted of A. The cytoplasmic membrane
To act as a physical barrier btw cytoplasm and environments and selectively controls the movement of substaces into and out of the cell“Semipermeable”
B. Cell wallThe rigid layer that protect the fragile cytoplasmic membrane from rupturingTo maintains cell’s shape
C. Capsule or slime layer (glycocalyx)
Cell Envelope
Cell membrane
PeripheralMembraneProtein
IntegralMembraneProtein
PeripheralMembraneProtein
Phospholipid
Bacterial plasma membrane are composed of 40 percent phospholipid and 60 percent protein. The phospholipids are amphoteric molecules with a polar hydrophilic glycerol "head" attached via an ester bond to two nonpolar hydrophobic fatty acid tails, which naturally form a bilayer in aqueous environments. Dispersed within the bilayer are various structural and enzymatic proteins which carry out most membrane functions.
Mesosome
The predominant functions of bacterial membranes are:
1. Osmotic or permeability barrier;
2. Location of transport systems for specific solutes (nutrients and ions);
3. Energy generating functions, involving respiratory and photosynthetic electron transport systems, establishment of proton motive force, and transmembranous, ATP-synthesizing ATPase;
4. Synthesis of membrane lipids (including lipopolysaccharide in Gram-negative cells);
5. Synthesis of murein (cell wall peptidoglycan);
6. Assembly and secretion of extracytoplasmic proteins;
7. Coordination of DNA replication and segregation with septum formation and cell division;
8. Chemotaxis (both motility per se and sensing functions);
9. Location of specialized enzyme system.
• Unique chemical structure– Distinguishes Gram positive from Gram-negative– bacteria and archaea bacterial species
• Rigidity of cell wall is due to peptidoglycan (PTG) – Compound found only in bacteria – Archaea –psudomurein or other sugars, proteins,
glycoproteins
• Many antimicrobial interfere with synthesis of PTG
• Penicillin; Lysozyme
Cell wall
• Basic structure of peptidoglycan– Alternating series of two
subunits• N-acetylglucosamin (NAG)• N-acetylmuramic acid (NAM)
– Joined subunits form glycan chain
• Glycan chains held together by string of four amino acids
– Tetrapeptide chain:L-ala-D-glu-DAP-D-ala L-ala-D-glu-Lys-D-ala
• Interpeptide bridge
Structure of peptidoglycan
Structures associated with gram-positive and gram-negative cell walls.
Differences of cell wall structure in Gram-positive and Gram negative cells
L FormsL Forms
Glycocalyx
CapsuleProtects bacteria from phagocytic cells
Slime layerEnable attachment and aggregation of bacterial cells
Capsules Most prokaryotes contain some sort of a polysaccharide layer outside of the cell wall polymer
Only capsule of B. anthracis consist of polypeptide (polyglutamic acid)
CapsuleCapsule
The capsule is covalently
bound to the cell wall.
Associated with virulence in bacteria.
Example:
Streptococcus pneumoniae
Slime LayerSlime Layer
The slime layer is loosely bound to the cell.
Carbohydrate rich material enhances adherence of cells on surfaces
Example:Streptococcus mutans and “plaque formation”
Biofilms Biofilms
The slime layer is associated with cell aggregation and the formation of biofilms
Example:Staphylococcus epidermidis biofilms on catheter tips
•Adhesion•Avoidance of immune response•Protection from dehydration •Protection of bacterial cells from engulfment by protozoa or white blood cells (phagocytes), or from attack by antimicrobial agents of plant or animal origin. •They provide virulent properties of bacteria (S. pneumoniae, B. anthracis)
General capsule function
Flagella
• 3 parts– filament – long, thin,
helical structure composed of proteins
– hook- curved sheath– basal body – stack of
rings firmly anchored in cell wall
• rotates 360o
• 1-2 or many distributed over entire cell
• functions in motility
Flagellar arrangements
1. Monotrichous – single flagellum at one end (cholera vibrio, blue pus bacillus),
2. Lophotrichous – small bunches arising from one end of cell (blue-green milk bacillus,Alcaligenes faecalis)
3. Amphitrichous – flagella at both ends of cell (Spirillum volutans),
4. Peritrichous – flagella dispersed over surface of cell, slowest E. coli, salmonellae of enteric fever and paratyphoids A and B
Bacterial MotilityBacterial Motility
The rotation of the flagella enables bacteria to be motile.
Flagella are important for:
Motility (dispersal)
Antigenic determinant
Number and location species specific
Pili and FimbriaePili and Fimbriae• Short, hair-like structures on the surfaces of procaryotic cells • Proteinaceuse filaments (~20 nm in diameter)• Very common in Gram-negative bacteria
• Functions:– Adherence to surface/ substrates: teeth, tissues– Involved in transfer of genetic information btw cells– Have nothing to do with bacterial movement (Except the twitching move
ment of Pseudomonas)
Fimbriae are smaller than flagella and are important for attachment
Bacterial endospores• Bacterial spores are often called “endospore” (since they are
formed within the vegetative cell)• Produced in response to nutrient limitation or extreme environments• Highly resistant• Highly dehydrated (15% water)• Metabolically inactive• Stable for years• Not reproductive • Functions: to survive under an extreme growth conditions such as
high temperature, drought, etc.
Bacillus, Clostridium, Sporolactobacillus, Thermoactinomyces, Sporosarcina, Desulfotomaculum species sporulate
Spore
Spores
• Key compositions:– Dipicolinic acid (DPA)– Calcium (Ca2+)
• Structure– Core / Cytoplasm– Plasma membrane– Core wall/ spore wall– Cortex – Spore coat– Exosporium
Endospores
The sporulation process begins when nutritional conditions become unfavorable, depletion of the nitrogen or carbon source (or both) being the most significant factor. Sporulation involves the production of many new structures, enzymes, and metabolites along with the disappearance of many vegetative cell components.
Spores are located: 1) Centrally (B. anthracis);
2) Terminally (С. tetani);
3) Subterminally (C. botulinum, C. perfringens)
The spores of certain bacilli are capable of withstanding boiling and high concentrations of disinfectants. They are killed in an autoclave exposed to saturated steam, at a temperature of 115-125 C, and also at a temperature of 150-170 C in a Pasteur hot-air oven.
Important Point: