culture media - prac. microbiology
TRANSCRIPT
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Bacteriological Culture Media
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Culture media provide optimal conditions
for growth & multiplication of bacteria.
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BACTERIOLOGICAL LOOP
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INOCULATION ON CULTURE PLATES
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According to the physical state media may be:
Fluid Media Solid Media
Growth is visualized as
Turbidity Colonies(macroscopic products of 20-30 cell divisions of a single bacterium on a solid medium)
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Plating-out technique
Procedure:
SpecimenSpecimen
Loopful of the specimen Loopful of the specimen
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Solid media are preferable:
Isolation in pure culture. Identification (colony morphology, effect on medium).
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Types of culture media:
1) Simple media
2) Enriched media
3) Selective media
4) Enrichment media
5) Indicator media
6) Selective and indicator media
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1- SIMPLE MEDIA
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1- SIMPLE MEDIA a- Nutrient broth
Meat Extract
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SEA WEEDS
Sea weeds
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AGAR POWDER
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1- SIMPLE MEDIA b- Nutrient agar
Nutrient Broth + 2% agar-agar
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1-SIMPLE MEDIAStaphylococcus aureus on nutrient agar (golden yellow endopigment)
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1- SIMPLE MEDIAPs. aeruginosa on nutrient agar (greenish exopigment)
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1-SIMPLE MEDIA Proteus on nutrient agar (swarming growth)
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2-ENRICHED MEDIA a- Blood agar ( also Indicator Medium)
Nutrient agar +5-10% Blood
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2- ENRICHED MEDIA Beta hemolysis on blood agar
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2- ENRICHED MEDIA Alpha hemolysis on blood agar
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2- ENRICHED MEDIA Non hemolytic organism on blood agar
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2- ENRICHED MEDIA Blood agar
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2- ENRICHED MEDIA b-Chocolate agar
Heated Blood agar
Haemoglobin
Heat
Haematin
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3-SELECTIVE MEDIA Lowenstein Jensen medium for M. tuberculosis
Beaten Eggs + Mineral salts + Malachite Green
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4- ENRICHMENT MEDIA Selenite F Broth
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5- Differential (indicator) media
Triple Sugar Iron (TSI) agar
TSI agar
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6- SELECTIVE INDICATOR (DIFFERENTIAL) MEDIA MacConkey’s medium
Bile Salts+
Lactose+
Neutral Red
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6- SELECTIVE INDICATOR (DIFFERENTIAL) MEDIA MacConkey’s medium
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Blood Culture
For isolation of bacteria from blood samples in cases of bacteraemia.
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Blood culture technique
(1)
(2)
(3)5 – 10 ml blood
Added to 50 – 100 ml fluid medium (broth)
Subculture on Solid medium
(4) Bacterial Growth
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Why use Large volume of broth???
1q111
1- Reduction of conc. of antibacterial constituents in blood
2- Multiplication of bacteria
3- Neutralization of antibiotics
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Anaerobic Cultivation
-For obligate anaerobes
Achieved by: - exclusion of oxygen
- addition of reducing substance
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ANAEROBIC MEDIAa- Robertson’s cooked meat medium
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ANAEROBIC MEDIA b- Thioglycollate Broth
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GAS PAK SYSTEM FOR ANAEROBIC CULTIVATION
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GAS PAK SYSTEM FOR ANAEROBIC CULTIVATION
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MCQs
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1) MacConkey’s medium:
a) Is an enriched medium.
b) Is an enrichment medium.
c) Contains phenol red as a pH indicator.
d) Gives pink colonies by lactose fermenting
bacteria.
e) Gives green colonies by lactose non-fermenting
bacteria.
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2) Blood culture technique is used to diagnose:
a) Sore throat
b) Conjunctivitis
c) Skin infection
d) Sinusitis
e) Bacteraemia
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3) All are correct about enrichment media except:
a)They are solid media.
b) Blood agar is not considered an enrichment medium.
c)They have selective properties.
d)They help multiplication of certain species and suppress others.
e)They include selenite broth.
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4) All are correct about chocolate agar EXCEPT:
a) It is an enrichment medium.
b) It contains heated blood.
c) It is used to grow fastidious organisms.
d) It is an enriched medium.
e) It is brown in colour.
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5) One of the following is used to grow strict anaerobes:
a) Selenite broth
b) MacConkey’s medium
c) Blood agar
d) Lowenstein-Jensen medium
e) Robertson’s cooked meat medium
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6) When strict anaerobes are grown in the Gaspak system:
a)They must be inoculated on cooked meat medium.
b)They can be grown on blood agar plates.
c) 50% carbon dioxide must be added.
d) The hydrogen released acts as a catalyst.
e)The palladium in the system acts as a nutrient.
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Thank You For Your Attention