Maseno University

School of Public Health and Community

Development

Department of Public Health

PHT 222:Medical Microbiology

 Medical Microbiology

At the end of this course, you will gain knowledge in the

following:
• Normal flora of the human body
• Microbiological disease states related to the human
body
• Media/Agar used to grow microorganisms
• Biochemical reactions of microorganisms
• Identification of microorganisms.
• Safety measures to prevent laboratory acquired
infections.
• Prevention and control of infections in the community.
 Medical Microbiology
• Microbiology – study of microorganisms, i.e.,
simple forms of life visible only through a
microscope and are usually single celled.
• Are found everywhere.
• Medical microbiology is the study of causative agents
of infectious diseases of humans and their reactions to
such infections. In other words it deals with etiology,
pathogenesis, laboratory diagnosis, specific treatment
and control of infection (immunization).
 Medical Microbiology
• When microorganisms are pathogenic in
nature or displaced from their natural
environment, they cause infections and
disease.
• You must understand different life forms of
microorganisms, how they may be identified,
and learn proper collection techniques for
common specimens.
 History
• Edward Jenner is credited with first vaccine – in
epidemics of smallpox during the late 1700’s he observed
that milk maids didn’t get the disease
• cattle had a similar disease – cowpox, milk maids had cow
pox lesions, but not small pox, he purposefully took
scrapings from cowpox blister and scraped a 8 year old
volunteer. With the material – child got mild illness but not
small pox,

• Vaccination comes from Latin word “vacca” meaning cow.

Jenner laid the foundation for Pasteur’s later work with
other vaccinations.
• (Vaccinia virus similar to Variola smallpox virus, today a
possible bioterror weapon – genetically engineered variola?)
 History
• Pasteur – French sceintist that dealt the death blow to the
spontaneous generation theory.

• He devised the ingenious curved necked flasks that

prevented contaminated air from reaching boiled beef broth
– the broth remained uncontaminated even though exposed
to the air

• He was very lucky – no endospores present, or it would

have failed
• (resistant to boiling)
 History
• 1. Pasteur developed a process we call Pasteuriztion – he
heated wine to kill contaminating microbes – cured sick
wine (today we heat treatment to kill pathogens in milk
also)
• 2. He proved that fermentation was caused by a microbe –
yeast
• 3. He developed vaccines for rabies and anthrax. Vaccines
led to immunity to diseases that routinely killed many
people, used to help people long before they understood
how they even worked (science of Immunology)
• 4. He began the revolution in science that led to the Golden
Age of Microbiology (from 1857-1914)
History – some of Pasteur’s tools of
trade
 History
• Robert Koch - Developed Koch’s postulates – important technique
for determining the actual microbial cause agent of a disease – more
later, German, contemporary of Pasteur, several very important
contributions
• 1. He discovered the tuberculosis bug (tubercle bacillus,
Mycobacterium tuberculosis)
• 2. He discovered the cause of anthrax (Bacillus anthracis) – from
blood of dead cattle, cultured bacteria in pure culture, injected
bacteria in live cattle and they died, then again cultured the bacteria in
pure culture. This led to the establishment of a procedure for
determining microbial cause of disease.
 History - Koch’s Postulates

1. The microbe must be present in

every case of the disease but
absent from healthy organisms.
2. The suspected microorganisms
must be isolated and grown in
pure culture.
3. The same disease must result
when the isolated microorganism
is inoculated into a healthy host.
4. The same micro-organisms must
be isolated again from the
diseased host.
 History

• Koch’s and Pasteur’s work helped establish

the “Germ Theory of Disease” – that
microorganisms cause disease (in people,
animals, and even plants)
 Modern medical microbiology
• Bacteriology – the science of bacteria, the causative
agents of a member of infectious diseases.
• Virology – the science of viruses, non-cellular living
systems, capable of causing infectious diseases in
man.
• Immunology – the science which concerned with
mechanisms of body protection against pathogenic
microorganisms and foreign cells and substances.
• Mycology – the study of fungi pathogenic for man.
• Protozoology – which deals with pathogenic
unicellular animal organisms.
 CLASSIFICATION OF MICROORGANISMS
Protista Vira
• DNA-viruses and RNA-viruses

• Eukaryotes Prokaryotes
• Fungi Blue-green algae
• Algae Bacteria
• Protozoa Scotobacteria Photobacteria
• Slime moulds 1. Class Bacteria

2. Class Rickettsias
3. Class Mollicutes
 Microbiological nomenclature
• In microbiology the binominal system of
nomenclature is accepted where each species
has a generic and a specific name. The generic
name is written with a capital letter, and the
specific name – with a small letter. For
example: the anthrax bacillus – Bacillus
anthracis; the tetanus bacillus – Clostridium
tetani.
 Method of Classification

• At present there is no universally accepted system

for bacterial classification.
• However, Bergey’s Manual of Determinative
Bacteriology is widely used.
• This approach uses a combination of fundamental
characters for bacterial classification
 Method of Classification cont….
• The characters considered during classification include:
– Morphology : Cell shape:
– Gram stain reaction
– Spore formation
– Biochemical reactions
– Nutritional requirements and
– Pathogenicity - Within one species, strains and subgroups can
differ by:
• the disease they produce
– Genetic relatedness (DNA composition)
• High G + C content
• Low G + C content
 Morphology

• Morphological

– Cell shape, size, colonial morphology,

mechanisms of mortility, staining behaviour,
cilia and flagelar, endospore shape and
location, spore morphology, cellular inclusions
and color.
Diversity of Bacterial Shapes and
Sizes
 Spherical
(cocci)
bacteria
1. Micrococci
2. Diplococci
3. Streptococci
4. Staphylococci
5. Tetracocci
6. Sarcine
 Representatives of pathogenic cocci

1 2

1.Scanning Electron Micrograph of Streptococcus pneumoniae

2.Scanning electron micrograph of a Staphylococcus aureus
Electron Micrograph of
Neisseria gonorrhoeae
 Rod-shaped bacteria
1
• Bacteria (1) include those
microorganisms, which, as rule, do
not produce spores (E. coli,
Salmonella spp, Shigella spp).
• Bacilli (2) (B. anthracis) and 2

clostridia (3) (C. tetani, C. botuli-

num) include organisms the majority
of which produce spores.
• Size of rod-shaped bacteria varies 2- 3

10 μm: small rods are 2-4 μm; long

rods are 5-10 μm.
ARRANGEMENT OF ROD-SHAPED
BACTERIA
Rod-shaped bacteria

1 2

1. Single Rod
2. Streptobacillus
 SPIRAL FORMS
• 1. Vibrios – are cells, which resemble a comma in
appearance (curved rods). Typical representative
of this group is Vibrio cholerae.
2. Spirilla – are coiled forms of bacteria. Pathogenic
species: Spirillum minus (1) – which is responsible for a
disease in humans transmitted through the bite of rats – rat-
bite fever – sodoku; Helicobacter pylori (2) – causative
agent of ulcer disease of stomach.

1 2
 SPIROCHAETES
• Treponema – exhibits, thin, flexible cells with 6-14
regular twists. The size of Treponema varies from
10-18 μ (T. pallidum).
• Leptospira – are characterized by very thin cell
structure. The leptospirae form 12-18 regular coils
(primary spirals) (L. interrogans) and C- or S- shape
according secondary twist.
• Borrelia – have large irregular spirals, the number of
which varies from 3 to 10. (B. recurrentis, B.
persica).
3. Spirochaetes – are flexuous spiral forms which include:
Treponema (T. pallidum) (1), Borrelia (B. recurrentis) (2),
Leptospira (L. interrogans) (3)

3
2
 Staining Reaction – Gram and
Acid Fast reaction
• Gram positive bacteria: Can retain Gram’s iodine in their
cell walls due to the amount of peptidoglycan in the cell
wall.
• Examples of genera are Bacillus, Listeria, Staphylococcus,
Streptococcus, Enterococcus, and Clostridium
• Gram negative bacteria: Can not retain Gram’s Iodine due
to less peptidoglycan in their cell walls.
– Examples are Escherichia coli, Salmonella, Shigella;
Pseudomonas, Moraxella, Helicobacter Stenotrophomonas,
Bdellovibrio, Neisseria gonorrhoeae, N. meningitidis.
 fuchsine

Gram
Staining
Technique
 Pathogenicity

• Within one species, strains and subgroups can

differ by:
– the disease they produce,
– their environmental habitat, site of infection and
mechanism of pathogenicity.
 Biochemical characteristics
• e.g. nutritional requirements.
• Lactose fermenters and non-lactose fermenters
• Catalase positive and catalase negative etc.
 Genetic Relatedness

• DNA composition

– High G + C content of DNA mol wgt - >50%.

– Low G + C content of DNA mol wgt < 50%.

• Can be achieved using similarity matrix or

dendogram tree.
BACTERIAL CELL
FLAGELLA

Electron Micrograph of Bacteria with Flagella

 CAPSULE

Enterobacter aerogenes Klebsiella pneumoniae

 SPORE

Bacillus anthracis Bacillus megaterium

 Cell wall
• In addition to conferring rigidity upon
bacteria, the cell wall protects against osmotic
damage
• Chemically, the rigid part of the cell wall is
peptidoglycan
• First described by Gram in 1884. It is' used to
study morphologic appearance of bacteria.
Gram's stain differentiates all bacteria into two
distinct groups:
• a. Gram-positive organisms
• b. Gram-negative organisms
 Bacteria with deficient cell walls
• Mycoplasma: a genus of naturally occurring bacteria
which lack cell walls
• L-forms: cell-wall-deficient forms of bacteria, usually
produced in the body of patients treated with penicillin
• Spheroplasts: derived from Gram-negative bacteria;
produced artificially by lysozyme or by growth with
penicillin or any other agent capable of breaking down
the peptidoglycan layer
• Protoplasts: derived from Gram-positive bacteria and
totally lacking cell walls; produced artificially by
lysozyme and hypertonic medium
Electron micrograph mycoplasma cells
(1) and L-forms of Proteus mirabilis
(2)

1 2
 Morphology of viruses
• Do not possess cellular organization
• Contain one type of nucleic acid
either RNA or DNA
• Lack enzymes necessary for protein
and nucleic acid synthesis
machinery of host cells
• They multiply by complex process
and not by binary fission.
• They are unaffected by antibiotics.
• They are sensitive to interferon.
Morphology of viruses
 Morphology of Rickettsiae.
• They are minute organisms having properties in
between bacteria and viruses.
• It contains both DNA and RNA.
• Contains enzymes for metabolic functions.
• Multiplies by binary fission.
• It is coccobacilli 300x600 nm in size, non-motile,
non-capsulated and is Gram-negative.
• Sensitive to many antibiotics.
• Can multiply only inside living cells.
Morphology of Rickettsiae
 Morphology of chlamydia.
• Chlamydiae are Gram-negative. They lack
some important mechanisms for the
production of metabolic energy, so they are
intracellular parasites. There are 2
morphological forms of chlamydia:
• Elementary bodies
• Initial bodies