Lab procedures when handling
micro-organisms
Distribution of micro-organisms in
nature
Learning Objectives
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Discuss the distribution of bacteria and fungi in
nature
Discuss and outline the Laboratory Procedures
for Micro-organisms
State precautions used when working with
micro-organisms
Define the terms: Asepsis & Sterility
Outline containment & disposal methods in
relation to microbes
Micro-organisms
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Can only be seen with the help of a microscope
Include bacteria, some fungi (yeast) and some
protists (plankton)
Occupy a wide range of habitats e.g. salt water,
fresh water, soil, dust, air, hot springs etc.
Fungi are mostly terrestrial
Bacteria can be found in extreme environments
from the inside of volcanoes to inside the human
gut
Laboratory procedures when
handling micro-organisms
Vocabulary
• Asepsis means that measures are taken
to exclude unwanted organisms
• Sterile means that all micro-organisms
are destroyed i.e. there is nothing living
Vocabulary
Inoculation is the addition of cells to the
nutrient medium
Incubation is the growing of the microbes
in a warm environment
Growth of micro-organisms
• Under suitable conditions one microorganism can be grown into a colony of
micro-organisms which is visible to the
naked eye.
• Micro-organisms are grown on a special
medium containing a food supply, most
commonly nutrient agar.
Growth of micro-organisms
• The procedures involved require caution as the
micro-organism to be grown (or any
contaminating micro-organism, if present) may
be a disease causing microbe.
• As a result certain precautions are taken when
handling micro-organisms to reduce the
possibility of contamination and to prevent the
growth of undesirable micro-organisms.
Precautions that can be taken
when handling micro-organisms
Wash hands before and after the experiment.
Precautions that can be taken
when handling micro-organisms
Wash the bench with disinfectant before and
after the experiment.
Precautions that can be taken
when handling micro-organisms
Sterilise all equipment before and after use. This
can be done by placing all equipment:
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In an autoclave (or pressure
cooker) at 120°C for 15 minutes
in Dettol for 24 hours
Precautions that can be taken
when handling micro-organisms
After an experiment material can be
placed in a dustbin only after sterilisation
Sterilised equipment can be reused
Precautions that can be taken
when handling micro-organisms
Flame all needles, loops and necks of testtubes by heating them in the flame of a
Bunsen burner.
Precautions that can be taken when
handling micro-organisms
• Turn off the Bunsen
burner when not in
use or make sure that
the flame is visible.
Precautions that can be taken when
handling micro-organisms
• Open all containers
for the shortest
possible time. Open
lids the shortest
possible distance.
Precautions that can be taken when
handling micro-organisms
Seal all plates once inoculated.
Precautions that can be taken
when handling micro-organisms
Label all plates once prepared.
Learning Check
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What is meant by asepsis?
What is meant by sterile?
State 3 precautions that should be taken
when handling micro-organisms.
State two ways in which equipment can be
sterilised.
How to grow a culture.
• Take a sterile agar
plate.
 Flame an inoculating
loop to sterilise it.
 Dip the loop into
sterile water or
alcohol to cool it.
•Flame the neck of the container from which the
sample is to be taken.
•Inoculate the loop by placing it briefly into the
sample to be grown.
•Re-flame the neck of the container.
While only slightly opening the agar plate
streak the inoculating loop across the surface
of the agar.
Sterilise the loop again by flaming it.
• Label and seal the
petri-dish and
incubate it upside
down for 2 - 3 days.
 Examine the plate
and record the
results.
Growth
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Once the plates are
sealed and labelled
turn them upside
down and place them
in an oven (incubator)
to allow any bacteria
and fungi to grow.
Placing them upside
down reduces
problems due to
condensation
Yeasts and moulds grow best at 25 - 30°C.
Bacteria generally require 35°C.
Examine the plate and record the results
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Bacterial colonies are
usually seen as white,
cream or yellow shiny
dots
Fungal colonies
appear as a powder
or as a fuzzy growth,
similar to cotton wool
in appearance
Disposal
All micro-organisms should be destroyed after
use by sterilising all petri-dishes and equipment:
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by placing them in an autoclave (or
pressure cooker) at 120°C for 15 minutes
or by placing them in Dettol for 24 hours.
Syllabus
Depth of treatment
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Precautions when working with microorganisms.
Asepsis and sterility: definition of each term
as applied to living organisms.
Containment and disposal.