Documente Academic
Documente Profesional
Documente Cultură
Sample
reception
@ Sample
cell
Acknowled
gement of
the sample
in the
system
(Ackn)
Sample
sent to the
lab for
analysis
with RoA
(Request of
analysis)
Sample
reception
in lab
Sample
preparation
Analysis is
continued
Analysis workflow
Confirmation
Plates marked for analysis
traceability
Sample
preparation
Result
Reading
Dilutions &
Plating
2.5 cm
Plates are stacked in a
stack of 6 petriplates .
Result
Recording
Salmonella
Shigella
Vibrio cholerae
Vibrio parahaemolyticus
Clostridium perfringens
Clostridium botulinum
Listeria monocytogenes
Legionella
All the parameters are analysed
using particular LI (Laboratory
Instructions) which are formulated
using ISO at NRC (Neste research
centre) .
In few cases NRC recommends to
ISO. (In theses cases we directly
follow the ISO)
9/16/2015
Non Pathogenic
CQAL Moga
Aseptic Technique
protective clothing
hand washing
bench cleaning
loop flaming
pipettors
Disinfecting workbenches
with 70% alcohol
Sterile Technique
Microbes in the laboratory can be found all around you, but
they are so small that you cannot see them. Microbes are in
the air, on benches and equipment and in water. They are one
of the major cause of the contamination of samples.
When culturing bacteria or other microorganisms, it is
important to keep your work area as clean as possible.
This prevents the introduction of other microorganisms from
the environment into your culture.
The techniques used to prevent contamination are referred to
as sterile or Aseptic techniques.
Sterile Technique
1. Start by washing your down your work
or lab benches with a surface
disinfectant. The most commonly used
disinfectants for lab use are:
1. 70% Ethanol
2. 200ppm Chlorine solution for
workbenches
3. 1000ppm chlorine solution for floors.
Microbial
Techniques
Line your sterile petri plates along the edge of the table. Transfer hot media to a
small sterile container and pour 15-20 ml of the plate media into each petri plate.
The petri plate lid should be open slightly, but not completely open as this
increases contamination. The thermal current created by the hot media prevents
bacteria and fungal spores from landing in your clean dish.
Store agar plates upside down. Stack the plates in their original bags for
further protection from contamination.
2. Store agar plates in a refrigerator. Most bacteria cannot grow well in cold
temperatures.
3. Store plates in a cold room if a refrigerator is not available. If you are storing
plates in a cold room, check the plates for condensation a few hours after
pouring. Condensation results from exposure to a heat source that drives
water out of the water and into the lid of the plate. This will dry the agar out
and render it unusable. Turn the plates over if condensation is visible and
monitor closely for more condensation development.
Before using the plates, examine them carefully for microbial growth (tiny colonies
of microbes) that may have grown during storage.
Check for cracking of the agar medium, which indicates that the plates are drying
out. If the plates are not dried out and have not been contaminated, the plates can
be used.
Stab Culture
By puncturing a
suitable medium with
a long, straight
charged wire.
For gelatin
liquefaction, stock
cultures & motility
Broth Culture
Inoculated by a
charged loop, pipette
or syringes.
For blood cultures &
sterility testing.
05.10.08
Dr Ekta, Microbiology
Quadrant streak
Flaming of the
loop Till it
becomes red hot
16 Streak
Stabbing( on the
butt) & streaking (on
the slat) is done on
agar slats
Dilution preparation
We get lesser
count as we
move to higher
dilutions
Counting colonies by
using colony counter
Controls
The samples are analysed by controlling all the
factors involved. To ensure this, four different
controls are set. These are:
AC: Agar control. Only the P+M media is
poured to ensure its sterility. If growth is
observed in this control, the media was
contaminated and hence this means the
growth obtained in other sample plates is not
solely of the sample but of the media too.