General Microbiology

Lecture 2

Dr/ Mohammed Ramadan

Lecturer of Microbiology & Immunology
 Cytoplasm
 Nucleoid
Chromosomes
• The bacterial chromosome exists as a singular, covalently closed circular
molecule of double stranded DNA.
• It is not associated with histones.
• The DNA is super coiled into a number of domains (c. 50) .
• Topoisomerases control topological changes in DNA architecture
Plasmids
• Plasmids are relatively small, circular pieces of double-stranded extrachromosomal DNA.
• They are capable of replication and encode functions that are not usually necessary for
bacterial growth.
• Increasing the spread of antibiotic resistance.
 II- Cytoplasm

Ribosomes:
present as tightly packed spherical particles in the
cytoplasm. Clusters of ribosomes are called polysomes.
- They are composed of 40% protein and 60% RNA.
- Bacterial ribosomes are 70S (Svedberg i.e. sedimentation
unit) in size, with two subunits 30S and 50S subunits.
- They are the site of protein synthesis.
- Eukaryote Vs prokaryote
-
 Mesosomes

They are invaginations of the cytoplasmic membrane.

Functions:

1- They play a role in respiration.

2- They play a role in cell division→

(septal mesosomes) where they function as:

a) The origin of the transverse septum that divides the cell in half

b) As the binding site of the cell DNA which duplicates to provide the genetic
material of each daughter cell.

3-lateral mesosomes increase surface area for transport.

 Flagella

Bacterial motility is commonly provided by flagella

• Flagellin - Hook - Flagellar motor
 Flagella

The numbers and distribution of flagella vary with bacterial

species.
1- Bacteria which have one flagellum at one pole are known as monotrichous.
2- Those with single flagellum at each pole are amphitrichous.
3- Bacteria with a bunch of "flagella at one or both poles are known as lophotrichous.
4- When the flagella are distributed all over the body of the cell, these bacteria are known as
peritrichous.
 Flagella
Various distributions of flagella
 Flagella

Functions:
1- Organ of locomotion.
a- Chemotaxis→movement of bacteria towards nutritives.
b- Aerotaxis→movement of bacteria towards optimal O2
concentration.
c- Phototaxis→movement of photosynthetic bacteria towards light.
2- Antigenic: H antigen is used for typing and diagnosis.
3- Penetrating through viscid mucus secretions and epithelial barriers
and spreading throughout body fluids and tissues.
 Fimbriae

Fimbriae
 Fimbriae are structurally similar to flagella, but are not
involved in motility.
 straighter, more numerous, thinner and shorter than flagella
 They are present in Gram-negative bacilli and responsible for
adhesion & attachment to mucosa. It acts as a virulence factor
of bacteria and composed of subunits of a protein, pilin
• Adhesions, allowing organisms to attach to surfaces of host.
• Haemagglutination and cell clumping in bacteria
Fimbriae
 Pili

• Pili are morphologically and chemically similar to fimbriae, but they

are present in much smaller numbers (< 10) and are usually longer.
• They are involved in the genetic exchange process of conjugation.
i) Ordinary pili: Also known as colonization antigens. These mediate
adherence of bacteria to specific receptors on human cell surface which is
a necessary step in initiation of infection of some organisms (e.g. mutants
of N. gonorrhea that do not have Pili are non pathogenic).
ii) Sex pili or F-pili: They are highly specialized hair-like structure found
in many bacteria ex E. coli and related bacteria. They play a role in the
transfer of part of the genetic material from one cell (F+ donor) to another
(F- recipient) thus giving rise to a simple form of sexual reproduction.
Types of Pili
 Differences between flagella and fimbria

  Fimbria Flagella

definition Thin, short thread like structure arise Long thin filaments arise from
from cell wall cytoplasm

function -adhesion -organ motility

-haemagglutination -serotyping

number More numerous less numerous

structure Pillin protein Flagellin protein

subtypes Absent Present

site Gram negative bacteria Both Gram negative& G+ve

Differences between fimbriae and pilli
Differences between fimbriae and pilli
 Capsules and Slime

 It is a thick gelatinous layer outside the cell wall of some bacterial

species.
 Many bacteria secrete extracellular polysaccharides (EPS) that are
associated with the exterior of the bacterial cell.
 The EPS is composed primarily of 2% carbohydrate and 98%
water,
 e.g. St. Pneumonia, N. meningitidis and H. influenza.
 But can be composed of other materials (e.g., polypeptide in B.
anthracis).
 N.B. Variation in the sugar components of the polysaccharide is
responsible for the presence of different.
Capsules
 Capsules and Slime

Forms of EPS:
Capsules form: a tight, rigid layer closely associated with the cell.
Slimes form: loosely layer associated with the cell.
Function of capsule and slime:
1- Protection against the penetration of biocides and disinfectants.
2- Act as a cement binding cells to each other in biofilms. Ex. alginate, produced by
Pseudomonas aeruginosa.
3- Outer polysaccharide layers in the attachment of certain pathogenic
microorganisms to their hosts.
4- Resist the phagocytosis
5- Identification of organisms due to the antigenic variation of the capsular material.
 Inclusion Granules
Inclusion can often be seen directly with the light microscope using special
staining procedure, but their contrast can be increased by using dyes. They
consist of:
1.Volutin (polyphosphate): known as metachromatic granules. These granules
are stained by many basic dyes; one of these dyes. Toluidine blue, become
reddish violet in color when combined with polyphosphate. It present in
diphtheria.
This phenomenon is called metachromacy (color change)
2. Lipid granules: polymerized poly B-hydroxy butyric acid PHB) e.g. in the
bacillus genus. This substance may act as a carbon and energy storage product.
3. Polysaccharide granules: Granules of either glycogen (red-brown) or starch
(blue) in the cytoplasm of certain bacteria when stained with iodine .
 Bacterial endospores
 Definition: They are small oval or spherical
cells which are highly resistant to unfavorable
conditions. They are known as endospores as
they are produced intracellularly.
 Spores are considered to be a dormant or
resting phase (no metabolic activity) of
bacterial cell that can remain dormant for
years.
 Bacterial endospores

 Main spore structure:

1- Nuclear material. 2- Minute amount of cytoplasm. 3- Spore
coat. 4- Spore cortex.
5- Exosporium (outer spore membrane) which contain
calcium &dipicolinic acid.
 Shapes of spore
 1. Central as C. botulinum
 2. Terminal as C. tetani
 3. Sub terminal C. perfringens
Bacterial endospores
Bacterial endospores
Bacterial endospores
Bacterial endospores
 Bacterial endospores

 Causes of marked resistance of spores:

1- Thick spore cortex and tough coat.
2- The bacterial spores contain large amount of
dipicolinic acid and calcium
3- Their dehydrated state.
4- Their very low metabolic and enzymatic
activity.
 Bacterial endospores

 Germination: Consist of 3 main steps:

1- Activation: On exposure to water and appropriate
nutrition.
2- Initiation: calcium dipicolinate is released and spore coat
and cortex are degraded by glycolytic enzymes.
3- Outgrowth: of new vegetative form that contains the
spore protoplast.
The germination is a rapid irreversible degradation
process.
 Bacterial endospores
 The medical importance of spores:
 Due to their extraordinary resistance to heat
and chemicals; Sterilization by autoclave at
121ºc for 20-30 minutes is required to
eliminate them (e.g., Geobacillus stearothermophilus).
 used as biological weapon as Bacillus
anthracis.
 Cell wall deficient bacteria

1. Protoplast, Spheroplasts, L. forms.

2. Mycoplasma.
They are deficient in cell wall. Hence, they are
pleomorphic and resistant to penicillin which
acts on the bacterial cell wall.
 Cell wall deficient bacteria

The process can be induced by:

1- Destruction of the cell wall by treating the organism with
an enzyme i.e. lysozyme which has a selective action on the
bacterial cell wall.
NB. In Gram negative bacteria the outer layer of the cell wall
is impermeable to lysozymes so must be disrupted with agent
as EDTA before treatment with lysozymes.
2- Cultivation of the organism in presence of penicillin which
inhibits the synthesis of cell-wall substance without
interfering with the growth of the organism.
 Cell wall deficient bacteria
3- They may be produced spontaneously.
 NB. If such treated cells are liberated from Gram
positive cells, they are called protoplasts (they must
be placed in isotonic solution to maintain the
spherical configuration and guard against cell lyses
and death because they entirely lack cell wall).
 NB. If such treated cells are liberated from Gram
negative cells they are called spheroblasts which
unlike protoplasts are osmotically insensitive because
they have intact cell wall lacking peptidoglycan only.
 Cell wall deficient bacteria

Spheroplasts Protoplasts
Cell wall lacking intact peptidoglycan Cell wall is completely removed
Arise from Gram negative bacilli by Arise from Gram positive organisms
growing it in the presence of by lysozyme hydrolysis of cell wall
penicillin
Osmotically insensitive Osmotically sensitive
Can multiply by binary fission Cannot multiply
Grow on ordinary media Grow on osmotically protective media
Can revert to parent form Cannot revert to parent form