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Culture methods ,
preparation and
sterilization of media
BACTERIOLOGICAL CULTURE DONE IN LAB:
• For isolating bacteria in pure culture from clinical samples
• To perform biochemical tests for identification of bacteria
• For antimicrobial sensitivity testing from isolating bacteria
• To maintain stock cultures
• To estimate viable bacterial count
LOOPS AND STRAIGHT WIRES
Made up of nichrome or platinum
 Bacteriological loops: 2-4mm internal diameter
used for streaking culture media
 Bacteriological straight wires: for making stroke
culture and stab culture
 Loops/wire is first heated in Bunsen flame
 by making it red hot and cool by waiting
10 sec.
 Done in biosafety cabinet to prevent lab acquired
infection
STREAK CULTURE
Routine method employed for isolation of bacteria or for
obtaining isolated colonies from mixed culture of bacteria.
LAWN OR CARPET CULTURE
It provides uniformly thick surface growth of the bacterium on
solid medium.
• Swabbing
• Flooding
USES:
1. For AST by disc diffuson method
2. Bacteriophage typing
3. Producing large amounts of bacteria growth required for
preparation of bacterial antigens and vaccines.
STROKE CULTURE
• For providing pure growth of
bacterium for carrying out
diagnostic tests
• In urease test
LIQUID CULTURE
Liquid cultures in test tubes,screw
capped bottles or flasks may be
inoculated by touching with a loop
by adding inoculum with pipettes or
syringes
USES:
1. Blood culture
2. Sterlity testing
3. Water analysis
INCUBATION OF CULTURE MEDIA
• At 37 degree celcius for 24 hrs in incubator.
• Candle jar- ex brucella abortus,
• streptococcus
• pneumococcus
• gonococcus
• Microaerophillic bacteria (campylobacter , helicobacter) require 5% oxygen
• In absence of oxygen
• By displacement and combustion of organism.
1. Mcintosh anf Filde’s anerobic jar
2. Anoxomat instrument
3. Gaspak system
4. Anerobic glove box
5. Anerobic work station
6. By using reducing age
-nts.
7. Prereduced anerobic
-ally sterlised.

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Culture methods ^J preparation and sterilization of media.pptx

  • 1. Culture methods , preparation and sterilization of media
  • 2. BACTERIOLOGICAL CULTURE DONE IN LAB: • For isolating bacteria in pure culture from clinical samples • To perform biochemical tests for identification of bacteria • For antimicrobial sensitivity testing from isolating bacteria • To maintain stock cultures • To estimate viable bacterial count
  • 3. LOOPS AND STRAIGHT WIRES Made up of nichrome or platinum  Bacteriological loops: 2-4mm internal diameter used for streaking culture media  Bacteriological straight wires: for making stroke culture and stab culture  Loops/wire is first heated in Bunsen flame  by making it red hot and cool by waiting 10 sec.  Done in biosafety cabinet to prevent lab acquired infection
  • 4. STREAK CULTURE Routine method employed for isolation of bacteria or for obtaining isolated colonies from mixed culture of bacteria.
  • 5. LAWN OR CARPET CULTURE It provides uniformly thick surface growth of the bacterium on solid medium. • Swabbing • Flooding USES: 1. For AST by disc diffuson method 2. Bacteriophage typing 3. Producing large amounts of bacteria growth required for preparation of bacterial antigens and vaccines.
  • 6. STROKE CULTURE • For providing pure growth of bacterium for carrying out diagnostic tests • In urease test
  • 7.
  • 8. LIQUID CULTURE Liquid cultures in test tubes,screw capped bottles or flasks may be inoculated by touching with a loop by adding inoculum with pipettes or syringes USES: 1. Blood culture 2. Sterlity testing 3. Water analysis
  • 9.
  • 10.
  • 11. INCUBATION OF CULTURE MEDIA • At 37 degree celcius for 24 hrs in incubator. • Candle jar- ex brucella abortus, • streptococcus • pneumococcus • gonococcus • Microaerophillic bacteria (campylobacter , helicobacter) require 5% oxygen
  • 12. • In absence of oxygen • By displacement and combustion of organism. 1. Mcintosh anf Filde’s anerobic jar 2. Anoxomat instrument 3. Gaspak system 4. Anerobic glove box 5. Anerobic work station 6. By using reducing age -nts. 7. Prereduced anerobic -ally sterlised.