INOCULATION ON CULTURE

MEDIA

By Dr Seemal Abbas
 INOCULATION

DEFINITION:
“ cultivation of bacteria on culture
media is called inoculation”
 INDICATIONS

• To isolate bacteria
• To demonstrate cultural characteristics
• To obtain sufficient growth for tets
• To determine antibiotic senstivity
• To maintain bacterial stock
 APPARATUS REQUIRED

•Petri dish
•Specimen
•burner
•Platinum wire
(straight wire: preferred for liquid media)
(wire loop: preferred to seed plate of
medium)
 PROCEDURE

1)STERILIZATION OF LOOP
2)SEEDING OF MEDIA
3)INCUBATION
1)STERILIZATION OF LOOP/NEEDLE

Hold the wire at the angle of 45 in flame untill it

becomes red-hot, allow it to cool
 2)SEEDING OF MEDIA

a)SEEDING OF LIQUID MEDIA (BROTH)

b)SEEDING OF CULTURE PLATE
 a)SEEDING OF LIQUID MEDIA (BROTH)
“used to increase initial conc. Of bacteria”

Take cooled sterilized wire >>>> first dip it in bacterial

suspension >>>> then in liquid media(broth)

Inoculating tube is incubated at desiured temp. for 24hours

Transparent broth turns turbid after inoculation,due to

bacterial growth
 b)SEEDING OF CULTURE PLATE
Take a loop full of specimen with cooled, sterilized wire loop
>>> gently streak in zig-zag pattern covering 1/3rd of plate

Rotate petridish clockwise 90* >>> with cooled sterilized

loop streak 3-4times through original streak

Again rotate petridish clockwise 90* >>> with cooled

sterilized loop streak 3-4times through 2nd streak

(streaks should be close together, but not overlapping)

SEEDING OF CULTURE PLATE
 3)INCUBATION
usually incubated at 37*C for 24 hours
 SMEAR PREPARATION
Put a drop of distilled water in the centre of glass slide

With cooled,sterilized wire loop pick-up colony from

plate, emulsify it in saline drop

Spread drop evenly on glass slide and air-dry it

Stain the slide and observe under microscope

 RESULT INTERPRETATION
Gram reaction: G+ve Gram reaction: G-ve
Morphology: G+ve cocci Morphology: rods/bacilli
Arrangement: Arrangement:
single, diplococci, groups single, diplobacilli
Number: moderate Number: moderate