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CULTURE MEDIA

Bursts the cells


Indicators
Tape
I. Culture Media Paper strips
Composed of nutrients used for growth and Bacteria pores
identification of microorganism Filtration
In Vitro o Culture media that cant be heated
o Inside to outside of the body o Membrane filter
In Vivo Cellulose Nitrate Filter
o Inside the body (growing) Pore diameter of 0.45
microns
II. Classification of Culture Media Remove bacteria
By Physical State Pore diameter og 0.2
o Solid media microns
1.5% - 2% nutrient agar Remove virus
o Semi solid media
0.5% - 1% nutrient agar IV. Preparation
o Liquid Planted Media (20mL per plate)
No agar needed o 2 plates (nutrient agar)
Nutrient Broth o 2 plates (blood agar)
By Use o 2 plates (chocolate agar)
o Simple Media Tubed Media (5mL per tube)
Non-fastidious o 2 Butt
Grows very fast o 2 Slant
Nutrient agar o 2 Butt Slant
o Enriched Media
Fastidious COMPUTATION:
Blood agar plate
Chocolate agar plate Each plate
Brain heart infusion media 20 mL x 6 plates=120 mL
o Enrichment media
Allow particular organisms
to be identified and Each tube
isolated 5 mL x 6 plates=30 mL
Selenite F salmonella
typhi
o Selective media Tubes + Plates
Inhibits growth of certain
bacteria
Permits the growth of 30 mL+120 mL+ 50 mL ( excess )=200 mL
others
Mannitol salt
staphylococci Agar = 28g in 1000mL of distilled water
o Differential media o dH20 = affects the pH of culture media (acidic)
Colonies of one kind of
organism to be 20 g
distinguished from x 200 mL=5.6 g of nutrient agar
another 1000 mL
McConkey agar lactose
fermenters (red clonies)
from non-lactose
fermenters (colorless) ASEPTIC TECHNIQUE

III. Sterilization I. Aseptic Technique


Successful cultivation of organism Pre-use sterilization of all instruments,
Avoid contamination vessels, media, etc. and their avoidance of
their subsequent contact with non-sterile
Heating
objects
o Most practical, efficient,
Spraying Tabletop
inexpensive means of sterilization
o Disinfectant
o Denatures enzymes
2.5% Hypochlorite
o Flaming
Filter air to remove cells
Dry heating
and spores
Mouth must be passed
Safety cabinet / Clean bench
briefly through the Bunsen
o Class II
burner
Prevent organism to fall Filtered air
into the vessel Open panel
Before and after closing o Class III
the vessel Gas tight
o Autoclaving Arm length
Moist heating For highly pathogenic
121 degrees C / 250 microorganisms
degrees F
Heat stable II. Vocabulary
15-20 mins Inoculum
15 psi o A small amount of material that
contains bacteria, viruses or other
microorganisms that is used to o Imprint method
start a culture Isolate microorganisms on
Culture the surface of the material
o Group of organisms obtained in a On the incubation, the
culture media distribution of the
o Mixed culture microorganisms on the
2 or more types of surface of the material
bacteria may be traced
o Pure culture o Direct plate method
1 type of bacteria Directly exposed to
Well isolated bacteria surrounding air
Colony For aerosol microbes
o Culture containing group of o Spread plate
bacteria forming on a solid culture Small volume of liquid
media inoculum is spread
o Isolated Surface of solid medium
Bacteria separated from Use of sterile bent rod
the group Give rise to lawn plate
Purpose o Flood Plate
To identify if it is Flooding surface of a solid
pure or medium with liquid
contaminated inoculums
Withdrawn excess
III. Loop and Needle inoculum with a sterile
Loop Needle Pasteur pipette
For Broth Thick cultured Give rise to lawn plate
Isolated colonies Butt slant test tube o Pour Plate
Inoculum on sterile empty
IV. Methods of Innoculation plate using sterile
Liquid Medium graduated pipette
Agar poured on the
o Loop or needle carrying the
inoculum
inoculum is dipped into liquid
Plate swirled gently to
medium, moved slightly and then
withdrawn distribute the inoculum
and agar
Solid Tube Medium
Solidify
o Streaking
Growth of microorganism
Inoculate surface of slants
on the surface and
Inoculating loop subsurface
Needle o Swab Inoculation
Swab Compact cotton wool
o Stabbing Sterile swab
One has to inoculate the Dry surface
deeper microaerobic or Tip is moistend
anaerobic portion of
o Sterile
butt/slope
Normal
Needle saline
Solid Plated Medium solution
o Clock streak (NSS)
thinning out the o Sterile
inoculum broth
Isolated colonies required medium
Inoculum is known to
contain large number of
cells