This document discusses various types of culture media used for growing bacteria. It provides details on the common constituents of culture media such as peptone, meat extract, and agar. It also describes different types of media including basal media, enriched media, differential media, and transport media. Various culture methods are outlined like streak plating, lawn culture, stab culture, and pour plating which allow isolating bacteria or assessing properties like motility.

1. Dr. Amit kumar
amitkumarshahi90@gmail.com

2. • Culture media:
Clinical
specimens eg
pus ,urine
Inoculat
e BACTERI
A Isolation
CULTURE
MEDIA

3. CONSTITUENTS OF CULTURE MEDIA
SODIUM
CHLORIDE AND
OTHERS
PEPTONE
MEAT
EXTRACT

4. • Peptone:
• source of nitrogen
• digested protein
• SOURCE: lean meat or other protein material (heart muscles , casein) and
digested by pepsin.
• Constituents: proteoses , aminoacids , inorganic salts (phosphates , potassium
and magnesium) and nicotinic acid and riboflavin.
• Neopaptone , proteose peptone , mycological peptone

5. • 0.5% sodium chloride : maintain the salt concentration of media
equal to cytoplasm of microorganism

6. • Agar /agar agar :
• solidifying agent (powder form)
• Melt in water after boiling
• Jellifies after cooling
• Seaweeds
• Components : long chain polysaccharides , protein , long chain fatty acids,
inorganic salt (calcium and magnesium)
• Concentration :
• Solid : 1-2%
• Semisolid : 0.5%
• 6% to inhibit proteus swarming

7. • Meat extract :
• Prepared : beef
• Constituent : protein degradation product , inorganic salts ,
carbohydrates and growth factors
• Yeast extract:
• Prepared : washed cells of Baker’s yeast
• Constituent : aminoacids , inorganic salts (potassium and phosphates) and
carbohydrates
• vitamins, carbohydrates, organic nitrogen compounds and salts.

8. • Blood and serum :
• Enriched media
• Fastidious bacteria
• 5-10% sheep blood
• Serum is sterilized after collection

12. I BASED ON CONSISTENCY

14. II) BASED ON INGRADIENTS
Peptone water

15. Peptone:1% peptone + Nacl (0.5%) + D/W
PEPTONE
NB

16. • USE OF BASAL MEDIA :
• preparation of enriched media
• NB (study bacterial growth curve)
• Study colony morphology
• Pigment demonstration
• Semisolid media: motility of bacteria ,
maintaining stock culture
NA +BLOOD
PSEUDOMONAS ON
NA

17. • B) complex media :
• other than basal media
• added with complex ingredients such as yeast extract or casein hydrolyse
• mixture of many chemical species in unknown properties.

18. C

19. D) SPECIAL MEDIA :
Loeffler’s serum slop Blood culture
media
i)

20. 5-10% SRBC + molten NA at 47o c = BA
Type of media : differential media and enriched
media
Blood
agar

21. • Chocolate agar: 5-10% SRBC + molten NA at 70o c = CA
• haemophilus influenza

22. – Loeffler serum slope: isolation of corynebacterium diphtheriae
Beef serum: solidifying agent
– +
– Heart muscle: amino acids
– +
– Peptic digest of animal tissue: amino acids
– +
• Sodium chloride: ions.
– +
– Dextrose: carbohydrate
– +
– Egg: solidifying agent

23. • Blood culture bottle: microorganism from blood (SEPTECIMEA , BACTERIMEA)
sodium polyanetholsulphonate (SPS) :
• inactivate serum factors toxic to organism
beef heart : amino acids
+
calf brain infusion : amino acids
+
disodium phosphate: buffer
+
sodium chloride : maintain salt = cytoplasm
+
glucose
•
• BRUCELLA
BHI AGAR + BHI
BROTH

24. • Ii) Enrichment media
• Liquid media + inhibitory substances
• used isolation of organisms from mixed culture (stool , sputum)
• Tetrathionate browth : salmonella typhae
• Selenite F broth : shigella
• Alkaline peptone water : vibrio cholerae

27. iii)

30. Magnesium Sulfate
Glycerol
L- Asparagine nitrogen and vitamins
Monopotassium
Phosphate
carbon source , favorable to the growth
of the human type tubercle
Prevent contaminants
coagulates
enhance organism growth
and act as buffer
enhance organism
growth and act as buffers

31. phenyl red
Citrate : bromothymol blue
Triple sugar iron : phenyl red
iv)

33. iv)

35. • CLED (Cysteine lactose electrolyte-deficient agar): differentiate LF
and NLF
• Inhibit swarming of proteus
• Specially use for urine specimen

36. v)

37. • Vi) Transport
medium
• Use for transporting of
samples
• Used for delicate
organism
• Eg : stuart’s medium :
Gonococci
• Buffer glycerol saline:
enteric bacilli

38. VII)

40. METHODS OF CULTURE
SPECIMENS CULTURE MEDIA

41. PLATINUM WIRE :
1) Bacteriological loop:
2) Bacteriological straight wire:
stroke
stab

43. Streak culture
Use : to obtained isolated
colonies

44. LAWN CULTURE/CARPET CULTURE
USE :
AST , BACTERIAL
TYPING

45. STROKE CULTURE
USE : URESE TEST
TO OBTAINED PURE
GROWTH

46. STAB CULTURE

47. POUR –PLATE
CULTURE AND SPREAD
METHOD
USE :
TO OBTAINED VIABLE COUNT
OF BACTERIA
1
ml
9ml nb +
1ml bs
9ml nb 9ml nb 9ml nb
1 ml 1 ml 1 ml

48. LIQUID CULTURE
DISADVANTAGE :
DONOT PROVIDE PURE CULTURE FROM MIXED GROWTH
IDENTIFICATION NOT POSSIBLE
EXAMPLE : BLOOD CULTURE
TURBIDITY

50. • Metal or glass jar with metal lid
with a screw (to close airtight)
• Pressure gauge and outlet and
inlet
• Air evacuate through outlet and
hydrogen induce through inlet
• Residual oxygen remove with
hydrogen gas
• sachet containing aluminium
pellets coated with palladium
react with hydrogen and replace
o2 in the form of water(h2o)
Chemical indicator : methylene blue

51. Gaspak system
sachet containing aluminium pellets coated with palladium
HYDROGEN TO FORM H2O (OXYGEN
REMOVED)
OXYGEN REMOVAL
sachet containing
aluminium pellets
coated with
palladium