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Classification of
Microorganisms
& Medium
Formulation

Classification of Microorganisms:
Microbes can be classified into four major groups:

1- Protozoa

2- Bacteria.

3- Fungi.

4- Viruses.

1- The Protozoa: These are unicellular organisms with protoplasm

differentiated into nucleus and cytoplasm.

Diameters in the range of 2-100 m.

The most important groups of medical protozoa are:

A-Amoeba: Entamoeba species. Mode of Motility: pseudopodia.

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B- Mastigophora: Mode of Motility: the Flagella.

Gastrointestinal flagellates: Giardia intestinalis
Urogenital flagellates: Trichomonas vaginalis
Tissue and blood flagellates: Trypanosoma, Leishmania .

Trypanosoma, and Leishmania .

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C- Ciliophora: motile by cilia.

Example: Balantidium coli.

D- Sporozoa: intracellular infection.

Example: Plasmodium that cause Malaria.

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2- The bacteria:

Bacteria are unicellular prokaryotic microorganisms that multiply by binary

fission.
Bacteria can be classified according to morphology, arrangement, and staining
reaction into the following groups:
1- Filamentous bacteria: Streptomyces: antibiotic producers.
2- True bacteria:
Cocci: Gram positive:
Gram negative:
Bacilli: Gram positive:
Gram negative:

Staphylococcus, Streptococcus.
Neisseria.
Bacillus, Clostridum, Corynebacterium.
Enterobacteriaceae, Brucella.

3- Spirochetes: Slender flexuous spiral bacteria.

Borrelia, Treponema, Leptospira.
4- Mycoplasma: The Smallest bacteria that lack of a rigid cell wall.
5- Rickettsiae and Chlamydiae: intracellular parasites.

3- The Fungi:

These are saprophytic or parasitic organisms possessing relatively rigid cell

walls.
Medical fungi can be divided into:
1- Mould: Branching filaments; hyphae, mycelium. Usually 2 to 10 m in width.
Example: Epidermophyton, Trichophyton,
Microsporum, Aspergillus.
2- True Yeasts: these are ovoid or spherical cells that reproduce asexually by
budding and sexually with formation of spores.
Example : Cryptococcus spp.
3- Dimorphic fungi:
Produce a vegetative mycelium in artificial
media, but are yeast like in infected lesions.
Example: Histoplasma.
4- Yeast- like fungi: Example: Candida ( Pseudomycelium).

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4- The viruses:

Viruses consist of DNA or RNA enclosed in a simple protein shell known as a

capsid.
General properties of viruses
They are very small in size, from 20-300 m.
They contain one kind of nucleic acid (RNA or DNA) as their genome.
They are metabolically inert
They are obligate intracellular parasites.
They are only seen by electron microscope.
Depend on the parasitized cell for survival and multiplication

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Structure of bacterial cells:

Size, Shape, and Arrangement of bacterial cells:
Morphology and arrangement of bacterial cells are criteria used for classification
of bacteria into following groups:
1. Cocci (Singular: coccus).
2. Rods (bacilli), (Singular: rod, bacillus).
3. Vibrios (Singular: vibrio).
4. Spirilla (singular :Spirillum)
5. Spirochetes. (Singular: Spirochaete).
1. Cocci:
These are round or oval bacteria measuring about 0.5-1.0 micrometer in
diameter. When they multiplying, cocci may form pairs, chains, or irregular
groups.

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Cocci in pairs are called diplococci, for example, meningococci

and gonococci.
Cocci in chains are called streptococci, for example Streptococcus pyogens.
Cocci in irregular groups are called Staphytococci, for example, Staphylococcus aureus.

2. Rods (bacilli):
These are stick-like bacteria with rounded, square, or swollen ends. They
measure 1-10 micrometer in length by 0.3-1.0 micrometer in width.
It may arranged in:
A- Chains, for example, Streptobacillus species.
B- Branching chains, for example, lactobacilli .
C- Mass together, for example, Mycobacterium leprae.
D- Remain attached at various angles resembling Chinese letters, for
example, Corynebacterium diphtheria.

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3-Vibrios:
These are small slightly curved rods measuring 3-4 micrometer
in length by 0.5 micrometers in width.
Most vibrios are motile with a single flagellum at one end.
They show a rapid darting motility.
For example:
vibrio cholerae.

4-Spirochetes:

These are flexible, coiled, motile organism, 6-20 micrometer in length.

They progress by rapid body movements.
Spirochetes are divided into three main groups:
A- Treponemes.

B- Borreliae.

C- Leptospires.

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Medium : a nutrient blend used to support microbial growth.

There are three physical forms of media, broth, solid, and

semisolid.

Solid media are more versatile in their usage.

Promote surface growth

Used to isolate pure cultures

Ideal for culture storage

Helpful in the observation of biochemical reactions

Used to make slants, deeps, and plates (named by

medium)
Mohammed L aqqan

A medium is sterilized (living organisms removed) before

usage in the lab.

Sterilization methods include; autoclaving, dry-heat,

filtration, UV exposure and ethylene oxide.

Culture: Is part of specimen grown in culture media.

Culture Media: is a medium (liquid or solid) that contains

nutrients to grow bacteria in vitro. Because sometimes we
cannot identify with microscopical examination directly, and
sometimes we do culture for antibiotic sensitivity testing.
Mohammed L aqqan

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Properties of Media:

Support the growth of the bacteria.

Should be nutritive (contains the required amount of
nutrients).
Suitable pH (neutral to slightly alkaline 7.3-7.4).
Suitable temperature, and suitable atmosphere.
(Bacteria grow at 370 C)
Note: media are sterilized by autoclaving at 1210C and 2
atmosphere for 15-20 minutes. With the autoclave, all bacteria,
fungi, viruses, and spores are destroyed. Some media cant be
sterilized by autoclaving because they contain eggs or
carbohydrates .
Mohammed L aqqan

Forms of culture

vSolid (agar): Is Broth plus agar (seaweed).

Are prepared by adding a solidifying agent (agar 1.5 3%).Prepared mainly in Petri dishes, but also in tubes
and slopes. After growth the bacterial colonies are
visible. e.g. blood agar, chocolate agar, MacConkey
agar.
vSemisolid agar (soft agar): Contains small amounts of
agar (0.5-0.7%). Used to check for motility and also
used as a transport media for fragile organisms. Can
have semisolid agar in Petri dishes or in tubes. In tubes
it is usually slanted to increase surface area.
Mohammed L aqqan

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Liquid (Broth): Mostly used for biochemical tests

(blood culture, Broth culture). Growth of bacteria is
shown by turbidity in medium. e.g. Nutrient broth,
Selenite F broth, alkaline peptone water.
Properties of agar:
Some what like gelatin.
It melts at 970 C and solidifies at 370 C.
Comes as sold powder and then you add water to it.
Mohammed L aqqan

Types of Culture Media

vSimple (basal, ordinary): Culture Media: are media that contain the basic
nutrients (growth factors) that support the growth of bacteria without special
nutrients, and they are used as basis of enriched media. E.g. Nutrient broth,
nutrient agar, peptone water. They are for the growth of non-fastidious
organisms like E. coli.

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Enriched Culture Media: are media that are enriched

with: Whole blood e.g. blood agar. Lysed blood (heated
to 80C) e.g. Chocolate agar.
Selective Media: it is a media, which contains
substances that prevent or slow the growth of
microorganisms other than the bacteria for which the
media is prepared for. For example
TSI (triple sugar iron agar): slanted tube.
EMB (Eosin Methylene blue): enteric isolation media.
Mohammed L aqqan

vDifferential Media (indicators): Contains indicators,

dyes, etc, to differentiate microorganisms. E.g.
MacConkey agar, which contains neutral red (pH
indicator) and is used to differentiate lactose fermenter
and non-lactose fermenter. (E.g. E. coli and
Salmonella).

Mohammed L aqqan

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Common media used in Microbiology Laboratory

Chocolate Agar: blood agar prepared by heating

blood to 95C until medium becomes brown or
chocolate in color heating the blood releases broth X
and V growth factors and also destroys the inhibitors
of V factor. These factors are required for the growth
of most species of Haemophilus and also Neisseria
gonorrhoear.
MacConkey Agar: an inhibitory and differential
medium used to distinguish lactose- fermenting Gramnegative organism from non fermentation. Crystal
violet, bile salts and neutral red are inhibitor agent.
neutral red is the PH indicator.
Mohammed L aqqan

vMannitol Salt Agar ( MSA ): for selective isolation for

coagulase positive, mannitol-fermenting staphylococcus.
Mannitol fermentation by pathogenic staphylococci is indicated
by a yellow halo surrounding the colonies.Sodium chloride is
the inhibitor agent.Phenol red is the PH indicator.
vMueller Hinton Agar: rich medium that support the growth of
most microorganism. It is commonly used for antibiotic
susceptibility testing: disk diffusion antibiotic susceptibility;
antibiotic serum level measurements; MBC determination.

Mohammed L aqqan

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vSalmonella Shigella ( SS ) Agar : isolation and differential

medium for pathogenic Gram-negative bacilli in particular,
Salmonella and Shigella. Inhibitor for Coliforms.
vTriple Sugar Iron Agar (TSI): this a key medium for use in
beginning the identification of a Gram- negative bacilli of the
enteric group. It contains glucose (0.1% ), Lactose (1%),
sucrose(1%). And peptone (2%) as nutritional sources. Sodium
Thiosulfate serves as the electron receptor for reduction of
sulfur and production of H2S. Detects fermentation of sucrose,
lactose, glucose, as well as production of hydrogen sulfide and
/or gas . Phenol red is the PH indicator; ferric ammonium
citrate is H2S indicator.

Mohammed L aqqan

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