Medical Microbiology

Cultivation of Microbes

Group #05
Syed Yawar Saeed, Haris Siraj,
Farooq Khan, Afaq Ahmad
BS 4th semester
Department of Microbiology
Abdul Wali Khan University Mardan
Cultivation of microorganisms
The process of growing microorganisms in culture by

taking bacteria from the infection site and grow them

in artificial environment in the laboratory .
Culture media
Culture Medium: Nutrients prepared for microbial
growth
 Inoculum: Suspension of microorganisms
 Inoculation: Introduction of microbes into culture
medium
 A pure culture contains only one species or strain
 Mixed culture contains several species
 Contaminated culture contains unwanted species of
organisms
Media

Classified according to three properties

Physical state
Chemical composition
Functional types
Physical state
Liquid Media
Liquid media are water based solutions that are
generally termed broths, milks, and infusions
Broth medium serves various purposes such as
propagation of large number of
organisms, fermentation studies,
and various other tests. e.g.
sugar fermentation test,
MR-VF broth
Physical state
Semi- solid media
Semi solid media contain a low percentage of agar
(less than 1%)
They have soft custard like
consistency and are useful
for the cultivation of
microaerophilic bacteria .
Physical state
Solid media
Solid media contain high percent of agar (1% - 5%)
It enables the formation of discrete colonies
Chemical content

Synthetic media :
Synthetic media contain pure organic or organic
compounds that are chemically defined “or”
A culture medium in which the amount and the purity
of all ingredients is known
Non Synthetic or enriched media:
Complex or enriched media contain ingredients that
are not chemically defined or pure i.e animal extract
etc
Functional Types of Growth Media
Basal media
An unsupplemented medium which promotes the
growth of many types of microorganisms which do
not require any special nutrient supplements
Functional Types of Growth Media

Enriched media
A complex medium to which “more” nutritious
ingredient is added &
Used to grow fastidious bacteria
Selective media
Supports the growth of some organisms but inhibits
the growth of others e.g Eosin Methylene Blue (EMB)
inhibits Gram-positives therefore selects for Gram-
negatives
Functional Types of Growth Media
Differential media
Differential or indicator media distinguish one
microorganism type from another growing on the same
medium.
 Eosin methylene blue is differential for lactose
fermentation
 MacConkey agar is differential for lactose
fermentation
 Mannitol salt agar is differential for mannitol
fermentation
Functional Types of Growth Media
Eosin Methylene Blue Agar
Functional Types of Growth Media
Manitol salt agar is a selective media
MacConkey agar is a differential media
Mannitol Salt Agar
Functional Types of Growth Media
Types of culture method (Streaking)
Types of culture method (Pour plate
& spread plate)
Environmental Requirements for Growth

Temperature
pH
Oxygen
Pressure
Temperature

Psychrophiles -optimum less than 20°C

Mesophiles-optimum 20-45°C
Thermophiles-optimum 45-80°C
Extreme Thermophiles-optimum 85+°C
pH

Acidophiles-optimum below pH 5.5

Neutrophiles-optimum at pH 6-8
Alkalophiles-optimum above pH 8
Molecular Oxygen (O2)

Microbe vary greatly in sensitivity to O2

Aerobes-microbes which require O2.
Anaerobes-microbes which DO NOT utilize O2AND
are KILLED by O2
Facultative Microbes-microbes which can grow in
presence OR absence of O2
Microaerophiles-required 3-15% O2
Molecular Oxygen (O2)

Microbe vary greatly in sensitivity to O2

Aerobes-microbes which require O2.
Anaerobes-microbes which DO NOT utilize O2AND
are killed by O2
Facultative Microbes-microbes which can grow in
presence OR absence of O2
Microaerophiles-required 3-15% O2
Molecular Oxygen (O2)

Aerobic Anaerobic Microaerophilic

culture culture culture
Pressure
Most organism on land and water are subjected to
pressure of 1 atm.
Hydrostatic pressure can reach from 600-1100 atm in
deep sea
Such as amphipods (Shewanella , Colwella)
Nutritional Requirements for Microorganisms

WATER
ENERGY
CARBON
ESSENTIAL ELEMENTS
Nutritional Requirements for Microorganisms

WATER-
-water serves as a solvent to carry nutrient to and waste
products away from the cell
- One method preservation of a microbial culture is
drying (desiccation).
Energy Sources

Phototroph-energy from sunlight

Chemotroph-energy from chemicals
- Chemoorganotrophs [organotrophs]-derive energy
from ORGANIC CHEMICALS
- Chemolithotrophs [lithotrophs]-derive energy from
INORGANIC CHEMICALS
Carbon Sources

Autotrophs-carbon from carbon dioxide

(CO2) -Inorganic carbon
Heterotrophs-carbon from Organic Carbon
Ex. Carbohydrates, lipids, protein
Essential Elements

 Hydrogen (H), sulfur (S), Oxygen (O), Phosphorus (P),

Nitrogen (N) commonly supplied as ammonia (NH4)
some microbes “fix” atmospheric nitrogen (N2)
Trace Elementsrequired in SMALL amounts Copper
(Cu), Zinc (Zn), Selenium (Se)
“Extra” Growth Factors

Fastidious Microorganisms
- microbes which need special organic chemicals added
to the medium for growth to occur
Ex. Vitamins, amino acids