GENERAL

MICROBIOLOGY

INTRODUCTION
TO
MICROORGANISMS

OUTCOMES OF GENERAL MICROBIOLOGY

By the end of the sessions, learners will be able to:
Differentiate among the major characteristics of each group of
microorganisms.
Identify the basic shapes of bacteria
Describe the structure and organization of bacteria
Compare and contrast the cell walls of gram-positive & gram negative
bacteria
Compare and contrast prokaryotic and eukaryotic cells
Describe Bacterial requirement for growth
Classify bacteria on the basis of preferred temperature range
Explain how microbes are classified on the basis of oxygen requirement
Describe Bacterial growth curve
Distinguish between different types of culture media
Identify bacterial viruses (bacteriophages)
Discuss bacteriophages propagation (replication)
Describe the mode of actions of antibacterial chemotherapeutic agents
Discuss antibiotic susceptibility testing
Identify the complications of chemotherapy
Describe the mechanisms of bacterial resistance to antimicrobial agents
Define Sterilization, Disinfection and antisepsis
Describe principles and applications of sterilization and disinfection
Define infection, specify the stages of the infectious process
Explain microbial virulence

Recommended books, periodicals

and web sites

Jawetz, Melnick, & Adelberg's Medical Microbiology Lang Medical Books/

McGraw-Hill (review of medical microbiology)
Greenwood et al. Medical Microbiology
Warren Livenson, Review of Medical microbiology & immunology
1. www.asm.org
2. www.virolgy.net/
3.www.pathmicro.med.sc.edu/book/wlcome.htm
4. www.youbundle.com/.../immunolgy-goldsby-immunology
5.http://www.biology.arizona.edu/immunology/microbiology_immunology.html
6. http://www.med.sc.edu:85/book/welcome.htm
7. http://www.mic.ki.se/Diseases/c2.html
8.http://www.tulane.edu/~dmsander/Big_Virology/BVHomePage.html
9. http://www.hepnet.com/hepb.htm
10. http://whyfiles.org/012mad_cow/7.html
11.http://www.microbelibrary.org/
12. asmnews@asmusa.org
13.http://www.phage.org/black09.htm
14.http://www.microbe.org/microbes/virus_or_bacterium.asp
15.http://www.bact.wisc.edu/Bact330/330Lecturettopics

OUT COMES of Bacterial Structure

By the end of the sessions, learners will be able
to:
Differentiate among the major characteristics of each
group of microorganisms.
Identify the basic shapes of bacteria
Describe the structure and organization of bacteria
Compare and contrast the cell walls of Gram-positive &
Gram negative bacteria
Compare and contrast prokaryotic and eukaryotic cells

Classification of Microorganisms
Microorganisms are either :
Prokaryotes e.g. bacteria
or Eukaryotes e.g. fungi

EUOKARYOTES
euo=true, karyote=nucleus
Algae, protozoa, fungi.
Cells of higher plants and animals
True nucleus, organelles, mitochondria,
Ribosomes (protein + RNA): 8O S (sedimentation

constant)
Divide by mitosis (& sexually)

PROKARYOTES
without true nucleus
Bacteria, mycoplasma, rickettsia, chlamydia

Hereditary material suspended in a portion of the

cytoplasm (not true nucleus)

No nuclear membrane
No organelles
Ribosomes: 70 S
No mitochonderia
Divide by simple binary fission

Prokaryotes

Eukaryotes

Size

1 10 um

10 100 um

Nuclear membrane

absent

present

Chromosomal No.

single

multiple

Ribosome

70 S

80 S

Peptido-glycan of
cell wall

present

absent

Replication

binary fission

Mitotic division

Membrane bound
organelles
Examples

absent

present

Bacteria

Fungi

BACTERIA:
THEIR STRUCTURES &
ORGANIZATION

Bacterial Morphology
Morphological features:

shape, size, arrangement, staining characteristics.

Bacterial size: 0.2-1.2 m width, 0.4-14 m length

Bacterial shape & arrangement:

(A) cocci (coccus): diplo, clusters, chains.
(B) bacilli (bacillus): single, in pairs or chains.
(C) Spirals: rigid or flexible.
Staining characteristics:

1. COCCI OR SPHERICAL

2. Bacilli or cylindrical:

3. Spiral:

Vibrio

Spirochetes

Staining characteristics:
kinds of stains: Simple, Differential.

Most important differential stain in clinical

microbiology is:

- Grams stain: it divides bacteria into

Gram+ve bacteria (violet staining) & Gram
ve bacteria (red staining).
- The 2nd important stain is Ziehl-Neelsen
stain, used to identify genus Mycobacteria.

BACTERIAL
ULTRA-STRUCTURE &

THEIR FUNCTIONS

Prokaryotic cell

Bacterial Structures
Bacterial Cytoplasmic
Coats
structures

External
Appendages

Capsule

Nucleoid material Flagella

Cell wall

Ribosome

Cell
membrane

Inclusion granules

Mesosome

Pili (Fimberia)

Cytoplasm
* It contains :
- Nucleoid :

Bacterial chromosome (1 mm long) single circular double stranded DNA

- Plasmids :
circular double stranded extrachromosomal DNA molecules
-

Ribosomes :
site of protein synthesis ( 70S )

Inclusion granules :
phosphates,sulpher,lipids,CHO---energy reserve, ATP synthesis

Mesosomes :
cell division, sporulation, respiratory enzymes

Cytoplasmic structures:
1- Nucleoid
- Single chromosome formed of single circular
molecule of ds DNA.
- Carries the genetic characters of the bacterium.
N.B.: Additional extrachromosomal Genetic
materials may be present in the
bacterium see later.

2- Ribosomes:
site of protein synthesis ( 70S )
3- Inclusion granules:
- These are granules of food & \ or energy
reserves.
- Phosphate (used for synthesis of ATP)
is stored in metachromatic or volutin granules.

4- Mesosomes:
- Composition:
inward invaginated convoluted tubules of the
cytoplasmic membrane into the cytoplasm.

- Functions:
Involved in the mechanisms of cell division and

sporulation.
Respiratory function: providing a membranous
support for respiratory enzymes.

Bacterial Coats
1- Cell membrane:
- Composition: Bilayer of phospholipoprotein (similar to that of
eukaryotic cells but without sterols)

- Functions of cell membrane:

1. Selective transport: simple diffusion, facilitated
diffusion and active transport.
2. Excretion of extracellular enzymes:
a- Hydrolytic enzymes: which digest large food
molecules
b- Destructive enzymes: used to destroy harmful
chemicals e.g. antibiotics.
3. Respiration: Due to presence of respiratory
enzymes
4. Reproduction.
5. Cell wall biosynthesis.
6. Chemotactic system

2- Cell wall:
- Composition:

- The cell wall of bacteria is a complex structure.

- Its strength is due to peptidoglycan [PG] (a complex

polymer consisting of N-acetylglucosamine [NAG]

and N-acetylmuramic acid [NAM]
- Besides PG, additional components in the cell
wall divide bacteria into Gram-positive and
Gram-negative.

- Gram-positive cell wall is composed of:

Peptidoglycan: There are as many as 40 sheets of PG,
comprising up to 50% of the cell wall material.
Teichoic acids:
- They are the polymer of ribitol or glycerol phosphate.
- Teichoic acids and cell wall associated
proteins are the major surface antigens of the
Gram-positive bacteria.

Gram-negative cell wall is composed of:

PG: It is much thinner, composed of only 1-2 sheets
comprising 5-10% of the cell wall material.
Outer membrane:
- It is phospholipid protein bilayer present external to the
PG.
- The outer surface of the lipid bilayer is composed of

molecules of Iipopolysaccharide (LPS) which consists

of a complex lipid called lipid A (endotoxin)
chemically linked to polysaccharides which represent
the major surface antigen of Gm-ve bacteria (somatic O
antigen).
Periplasmic space: It is the space between the
cytoplasmic and outer membranes. It contains the PG
layer and a gel-like solution of proteins.

Cell Wall: Gram-Negative & Gram-Positive

 Functions of the cell wall

1. It maintains the characteristic shape of the
bacterium.
2. It supports the weak cytoplasmic membrane
against the high internal osmotic pressure.
3. It plays an important role in cell division.
4. It is responsible for the staining affinity of the
organism.
5. It protects the organism from external
environmental hazards.

Wall deficient variants

(Bacteria without cell wall)

Mycoplasma

L Forms

Naturally present

Artificially induced due to

treatment of the cell wall by
antibiotics or lysosyms
Can replicate
Cant replicate
Cant reassume a May (or may not) reassume the
walled form
walled form upon removal of
the causative agents.
- Have no defined recognizable shape.
- Not stained by gram stain.
- Resistant to wall inhibitor antibiotics.

3- Capsule and Related Structures:

- Composition:
The outer most layer in some bacteria that is
formed only inside the host (In vivo)
It is formed of polysaccharide (except B.
anthracis capsule formed of polypeptide).

Functions:
A- Protection:
* Against phagocytosis (virulence factor).
* Against various kinds of antibacterial agents,
e.g. bacteriophages, colicins, complement
and lysozymes.
B- Attachment to host surface:
By capsules or glycocalyx in order to
establish infection, e.g. Streptococcus mutans
form glycocalyx to stick to the tooth enamel.

Capsulated bacteria

- Variants of the capsule:

1- Capsule:
- A well defined layer that adhere to the
bacterium surface all-around.
- It may be in the form of large capsule, small
capsule or microcapsule (electron dense
layer).
2- Slime layer: It is a surface layer that is loosely
distributed all-around the bacterium.
3- Glycocalyx: It is a loose meshwork of
polysaccharide fibrils extending outwards from
one pole of the bacterium.
N.B.: All capsule variants are not stained by Gram stain
and appear as hallow zones around the organism.

External appendages:
1- Flagellum:
- Composition: long tubular external appendages which
is formed of protein called flagellin (antigenic protein
and form the H antigen of bacteria)
- Function: flagellum is the organ of motility of the
bacteria [motility chemotaxis may be toward (+ve) or
away (-ve) from certain area]

Location and number of flagella

Flagella arrangement
A. Monotrichous: One flagellum
B. Lophotrichous: multiple polar flagella
C. Amphitrichous: a single flagellum on
each of two opposite ends

D. Peritrichous: Flagellae around the cell

E.M. picture of flagellated bacilli

2- Pili or fimbriae:
- Composition:
Thin short tubular external appendages which are
formed of protein called pillin (non antigenic protein).

- Types and Functions:

A- long pilus: also called the sex pilus (F or fertile pilus)
is involved in the transfer of DNA between bacteria, a
process known as conjugation.
B- Short pilus: also called fimberia and it adheres the
organism to the host surface (a virulence factor).

Pili (fimbriae)

Comparison between flagella and

pili
Morphology
Composition
Antigenicity
Functions

Flagella

Pili

Tall, thin &

tubular
Flagellin
protein
Antigenic

Short, thin &

tubular
Pillin protein

Motility of the
organism

-Attachment to
the host
- Gene transfer
by conjugation

Not antigenic

Bacterial Spores(Endo-spores)
- Definition: a highly resistant resting form (phase) of
some bacteria (genera Bacillus and Clostridium) that is
formed on exposure to bad external environmental
conditions.

- Sporulation: it is a process by which a single

vegetative bacterium forms a single spore on exposure

to bad external environmental conditions ( in vitro).

N.B.:Endospore does not grow or reproduce, and

exhibits absolute dormancy.

 Mechanism of sporulation:
- The cell membrane invaginates enclosing a section of the
cytoplasm that contains the bacterial chromosome, some
ribosomes and other cytoplasmic materials that will be needed
for germination.
- It acquires a thick cortex and a thin but tough outer spore coat.

- Germination: It is the process by which the

endospores can respond quickly to changes in the
environment returning to the vegetative state
within 15 min.
- Mechanism of germination: In the process of
germination, the spores absorb water and swell, the
protective coat disintegrates and a single vegetative
cell emerges.

Viability and resistance of the spore:

- The spores are highly resistant to disinfectants,
drying and heating and only moist heat at 121 C for
10 -20 minutes is able to kill spores.
- The marked resistance of the spores is due to:
1- High content of Ca 2+ and dipicolinic acid
2- The impermeability of their cortex and outer coat.
3- Their low content of water.
4- Their very low metabolic and enzymatic activity.

Morphological characters of endospores:

1. Staining: Using Gram's stain, the spore remains
uncoloured and can be seen as a clear area within
the stained cell. The spores can be stained using
special procedures.
2. The position: In relation to the body of the
bacillus, the spore may be central, terminal or
subterminal.
3. The shape: The spores may be oval or rounded.
N.B.: The position and shape of spores are
characteristic of the species and may help in the
microscopic identification of the bacterium.

Characteristics of Spores
Morphology
Staining
Position

Comparison between bacterial capsule

and bacterial endo spore
Capsule

Spore

Site of
formation

In vivo (inside the

host)

In vitro (in the

environment

Metabolic
activity and
replication
Functions

The organism is
active and can
replicate
Adhesion to the
host, protection
and antiphagocytic

The organism is
inactive and
cant replicate
Protection from
bad conditions
of the
environment