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Bacteriology
Lecture 1 د. محمد الفحام
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Microbial cell structure
A major biologic division separate the eukaryotes organisms containing a
membrane bound nucleus from prokaryotic organisms in which DNA is
not physically separated from the cytoplasm (Primitive cells).
Eukaryotic are termed protists which include algae, protozoa, fungi and
slime molds. They are large in size (true cells).
Prokaryotic include bacteria
mycoplasma, chlamydia and rickettsia.
Eukaryotes and prokaryotes are organisms because they contain all of
enzymes required for the replication and posses the biologic equipment
necessary for the production of metabolic energy so they are
distinguished form viruses which depends upon host cells for there
necessary functions.
Optical method:
Are methods for observation and study of microorganisms:
1. The light microscope: the resolving power of light microscope (L.M.)
is about half the wave length of the light being used. Microscopes employ
a 90-power objective lens x10 power ocular lens that magnifying the
specimen 900 times.
2. The electron microscope: have a high resolving power that is used to
observe the detailed structures of prokar. and eukar cells. This is due to
the fact that electrons have a much shorter wave length than the photons
of white light.
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EM is of two types:
a. Transmission electron M. (TEM)
b. Scanning electron M. (SEM).
3. Dark field illumination: There is a special condenser is used that both
blocks direct light rays and reflects it off a mirror on the side of the
condenser at an oblique angle. Its used for observing organisms (such as
spirochetes) which is less than 0.2μm in diameter and therefore cannot be
observed with direct light.
4. Phase phase microscopes used for examination of internal structures in
transparent living cells using a special optical systems.
5. Autoradiography: is used to reveal the position of the radio active label
cells that have incorporated radio active atoms such ah in cases in
following the replication of DNA.
Prokaryotic
The prokaryotic cells simpler than the eukaryotic cell at every level, with
one exception: the cell envelope is more complex.
Bacterial cell organelles
The Nucleoid:
1. It’s equivalent to the eukaryotic nucleus can be seen in L.M. it feul-
gen-positive indicating the presence of DNA. A histone-like proteins
exist in bacteria DNA similar to histone protein that bind to eukoryotic
DNA by ionic interactions.
2. The nuclear membrane and the mitotic apparatus present in eukaryotic
cells are absent in prokaryotes.
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3. The nuclear region is filled with DNA fibrils the nucleoid of bacterial
cells consist of a single continuous circular folded molecules of haploid
chromosome.
4. The number of copies of these chromosomes in a cell depend on the
stage in the cell cycle.
5. How ever bacteria have a liner chromosome as in Borrelia Bugdorferi.
6. A small amounts of RNA are present in nucleoid of most bacteria.
7. Bacterial DNA is associated at one point with an invagination of the
cytoplasmic membrane called a mesosome this attachment plays a role in
the separation of the two sister chromosomes following replication.
8. a. on the other hand the nucleus of eukaryotic cell is bounded by a
membrane continuous with the endoplasm reticulum. The nuclear
membrane exhibits the selection permeability between nucleus and
cytoplasm.
c. Eukaryotic chromosome contain linear DNA arrange in double helix in
the nucleus.
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3. The nuclear region is filled with DNA fibrils the nucleoid of bacterial
cells consist of a single continuous circular folded molecules of haploid
chromosome.
4. The number of copies of these chromosomes in a cell depend on the
stage in the cell cycle.
5. How ever bacteria have a liner chromosome as in Borrelia Bugdorferi.
6. A small amounts of RNA are present in nucleoid of most bacteria.
7. Bacterial DNA is associated at one point with an invagination of the
cytoplasmic membrane called a mesosome this attachment plays a role in
the separation of the two sister chromosomes following replication.
8. a. on the other hand the nucleus of eukaryotic cell is bounded by a
membrane continuous with the endoplasm reticulum. The nuclear
membrane exhibits the selection permeability between nucleus and
cytoplasm.
c. Eukaryotic chromosome contain linear DNA arrange in double helix in
the nucleus.
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Lecture 2 د. محمد الفحام
Cytoplasmic structures:
1. Prokaryotic cells lack the endoplasmic reticulum and the self
reproducing plastid as mitochondria, golgi apparatus and chloroplasts that
present in eukaryotes.
2. The electron transport enzyme for respiration localized in the
cytoplasmic membrane instead of mitochondria.
3. The photosynthetic pigment of photosynthetic bacteria are localized in
membrane arrangement vesicles underlying the cell membrane.
4. Bacteria often store reserve material in the form of insoluble granules.
When the source of nitrogen, sulfur or phosphorus is limited or when the
pH is low, excess carbon in the medium is converted to various polymers
of glucose such as starch and glycogen. These granular are then used as
carbon source when protein nucleic acid synthesis is resume. Similarly
certain bacteria oxidize the sulfide form H
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S producing intracellular
granules of polyphosphate as reserves of inorganic phosphate granules
are termed volutine granules or metachromic granules that are present in
corynebacteria.
5. Microtubular structures which are characteristic of eukaryotes are
generally absent in prokaryotes.
6. Certain groups of bacteria contain protein-bounded vesicles in their
cytoplasm.
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The Cell Envelope
The layers that surround prokaryotic cell are referred collectively the cell
envelops. The bacteria are differentiated to gram-positive and gram-
negative bacteria according to the difference in their cell envelope.
The G+ve envelope is relatively simple consisting of two-three layers:
1. The cytoplasmic membrane.
2. A thick peptidoglycan layer.
3. In some bacteria an outer layer called capsule
The G-ve envelope is highly complex multilayer structure:
1. Cytoplasmic membrane (inner membrane in G-ve-).
2. Single planar sheet of petidoglycan.
3. Complex layer called outer membrane.
4. An outer most membrane capsule may be present.
• The space between inner and outer membrane is called periplasmic
space.
The cytoplasmic membrane
Structure:
The cell membrane is a typical “unite membrane” composed of
phospholipid and proteins.
The membrane of prokaryotic differ from those of eukaryotic by the
absence of sterols. Convoluted invaginations of cell membrane (C.M.)
specialized structures are called “mesosomes”.
There are two types of mesosomes:
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1. Septal mesosomes that function in the formation of cross walls during
cell division. The bacteria chromosome (DNA) is attached to septal
mesosomes.
2. Laterla mesosomes.
Function of Cytoplasmic Membrane:
1. Selective permeability and transport of solutes: the C.M. forms a
hydrophopic barrier impermeable to most hydrophilic molecules. Several
transport systems are exist that enable the cell to transport nutrients into
and waste product out of the cell. These transport systems work against a
concentration gradient and increase the conc. of nutrients inside the cell a
function that requires energy.
There are 4 general transport mechanisms involved in membrane
transport:
a. Facilitated diffusion.
b. Binding protein-dependent transport.
c. Group translocation.
d. Chemiosmotic driven transport.
2. Electron transport and oxidative phosphorylation. The cytochrome and
other enzymes of the respiratory chain are located in the cell membrane in
prokaryotes is a functional analog of the mitochondrial membrane in
eukaryotic cells.
3. Excretion of hydrolytic exoenzymes that degrade the organic polymers
to subunits small enough to penetrate the cell membrane. All organisms
that use macromolecular organic polymers (as proteins, polysaccharides
and lipids) as a source of nutrients excrete hydrolytic enzymes.
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4. Biosynthetic functions: cell membrane is the site of the carrier lipids
and enzymes used in cell wall synthesis also it carry some proteins of the
DNA replication complex.
5. Chemotactic systems: organs of locomotion of bacteria as flagella bind
to specific receptors in cell membrane.
The cell Well
Functions:
1. The osmotic pressure of bacteria is sufficient to burst the cell unless the
presence of the strong cell wall that composed of peptidoglycan (murein)
(or it's called mucopeptide).
2. Bacteria are classified to G+ve or G-ve according to their response to
the differential stains method called Gram’s stains. The difference reside
on the cell wall differences.
3. Cell wall plays an essential role in cell division as well as its own
biosynthesis.
4. Cell wall is the site of antigenic determinates of the cell surface as the
lipopolysaccaride of G-ve cell wall that represents the endotoxin activity
of such bacteria.
5. Cell wall that selectively permeable.
Structure: The basal structure of cell wall is the peptidoglycan layer.
1. The peptidoglycan layer: it is a complex polymer consist of three parts:
a. A backbone composed of alternating N-acetyl glucosamine and N-
acetyl muramic acid.
b. A set of identical tetrapeptide side chains attached to N-acetyl muramic
acid.
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c. A set of identical peptide cross bridges,
the tetrapeptide side chain have:
- L-alanine at position 1
- D-glutamate at = 2
- - Diaminopimelic acid at = 3 or L-lysine in some G+ve bact.
- D-alanine at = 4
2. Special components of G+ve cell wall contain a thick layer of
peptidoglycan and teichoic acid and teichuronic acid that account for 50%
of cell wall weight. In addition some G+ve wall may contain
polysaccharide. There are 2 types of teichoic acid:
a. Wall teichoic acid that linked to peptidoglycans.
b. Membrane teichoic acid that linked to membrane glycolipid. Teichoic
acid constitute the surface antigens of G+ve bact.
3. Special components of G-ve cell wall: three component lie outside a
single planner sheet of peptidoglycan layer, these component are:
a. Lipoprotein: that cross link the outer membrane and peptidoglycan
layer and stabilize them together.
b. Outer membrane: it’s a bilayered structure: and inner leaflet resembles
in composition the cell membrane. While the phospholipids of the outer
leaflet are replaced by lipopolysaccharides molecules. Thus this bilayer
differ from the bilayer of cell membrane. The outer membrane protect the
cell from hydrophobic as well as hydrophilic molecules. The outer
membrane has special channels called porins that permit the passive
diffusion of low molecular weight hydrophilic compounds as sugars and
amino acid also large antibiotic molecules penetrate the outer membrane
slowly that makes G-ve bacteria much resistant to antibiotics.
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c. Lipopolysaccharide: it consist of complex lipid called lipid A to which
is attached a polysaccharides molecules that LPS is extremely toxic to
animals has been called endotoxin of G-ve. It’s firmly bound to the cell
surface and is released only when the cells are lysed. When LPS is split
into lipid A and polyssacharide all of toxicity is associated with the
former. (lipid A). The polysaccharide on the other hand represents a
major surface antigen of bacteria called O-antigen.
d. The periplasmic space: the space between the inner and the outer
membrane called the periplasmic space which contain the murein layer
(peptidoglycan) and a gel like solution of proteins. The periplasmic
proteins include binding proteins for amino acids, vitamins, sugars
hydrolytic enzymes and detoxifying enzymes that inactivate certain
antibiotics.
Cell wall growth:
As the protoplast increase in mass, the cell wall is elongated by the
intercalation of newly synthesized sub-units into the various wall layers.
Protoplasts, spheroplasts and L-Forms:
Removal of bacterial wall may be accomplished by hydrolysis with
lysozymes or blocking peptidoglycan biosynthesis with an antibiotic such
as penicillin. In osmotically protective media such treatment librate
protoplast from G+ve bacteria and spheroplast from G-ve bacteria (retain
outer membrane and entrapped peptidoglycans). If such cells are able to
grow and divide they are called L-forms. L-forms are produced with
penicillin treatment rather than lysozyme since they need the presence
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residual peptidoglycan. Some L-Form can revert the normal form upon
removal of inducing stimulus. Thus they are able to resynthesize a normal
cell wall from the residual peptidoglycan some bacteria produce L-Forms
spontaneously. L-Forms produce chronic infection in the host since they
persist in protective regions in the body. Since L-Forms do not posses a
cell wall so L-Forms infections are resistant to antibiotic as penicillin that
act on the cell wall of bacteria.
