Microbial Culture Media Composition Part-2

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Continued from Microbial Culture Media Composition Part-1

Some important media composition are given below.

11. Hugh and Leifson's medium
Peptone -0.2 gm
NaCl -0.5 gm
K2HPO4 -0.03 gm
Agar-agar -0.3 gm
Distill water -100 ml
0.2 %  Bromothymol blue -15 ml
Glucose -1 gm
Paraffin oil -10 to 20 ml
pH -7.1


It is used in oxidative – fermentative test .

a) Peptone is a source of carbon,nitrogen and energy.
b) NaCl maintain osmotic balance.
c) K2HPO4   acts as a buffer.
d) Agar is a solidifying agent.
e)  Bromothymol blue  is a indicator dye.
f) Glucose is a source of carbon.
g) Paraffin oil is used to maintain anaerobic condition.

Note – Bromothymol blue  is a indicator dye in acidic condition it shows yellow colour where as in neutral condition it shows blue colour  and     in alkaline pH it shows green colour.

12. Amminoacid decarboxylation media
Peptone -0.5 gm
Meat extract -0.5 gm
Glucose -0.05 gm
Pyridoxin hydrochloride -0.5 gm
Distill water -100 ml
pH -6.7
Bromocersol purple -(1: 500)  5 ml
Cersol red -2.5 ml
L-lysin( Monohydrochloride)- 1 gm

Role – It is used for isolation of different genera of species Enterobacteriace.

a) Meat extract acts as a source of mineral growth factor and amino acid.
b) Glucose is a source of carbon and it produces acid plus gas after fermentation.
c) Pyridoxine hydro-chloride is a co-factor it enhances the rate of reaction.
d) Bromocersol purple and Cersol red  are indicator dye.
e)  L-lysin is a test compound.

13. Peptone nitrate broth
Peptone -0.5 gm
KNO3 -0.2 gm
Distill water -100 ml
pH -7

Role – It is used peptone nitrate reduction test.

a) Peptone is a source of carbon, nitrogen and energy.
b) KNO3   is  a test compound.

14. Arginine broth
Tryptone -0.5 gm
Yeast extract -0.5 gm
Na2HPO4 -0.2 gm
L-arginine -0.3 gm
Dextrose -0.05 gm
Distilled water -100 gm
pH -7

Role – It is used in Arginine hydrolysis test.

a) Tryptone is a source of carbon,nitrogen and energy.
b) Yeast extract acts as a mineral,growth factor and amino acid.
c) Na2HPO4  is a buffer.
d) L-arginine is a monochloride and a test compound.
e) Dextrose is a source of carbon and on fermentation it produces acid and gas.

15. Phenyl alanine deamination medium.
Yeast extract -0.3 gm
L- phenyl alanine -1 gm
Na2HPO4 – 0.1 gm
NaCl -0.5 gm
Distill water -100 ml
pH -7.2
Agar -agar -3 gm

Role – It is used in Phenyl alanine deamination test.

a) Yeast extract provide luxuriant growth of bacteria as it is a source of growth factor, and amino acid.
b)  L- phenyl alanine  is a test compound.
c) Na2HPO4  is a buffer.

16. Deoxycholate citrate agar
A) Solution A
Sodium citrate -17 gm
Sodium thiosulfate -17 gm
Ferric citrate -2 gm
Distill water -100 ml
B) Solution B
Sodium deoxycholate -10 gm
Distill water -100 ml
C) Complete medium
Nutrient agar -100 ml
Lactose -1 gm
Neutral red -0.002 gm
Solution A -5 ml
Solution B -5 ml
pH -7.4

Role – It is used for isolation of Shigella and Salmonella.

a) Sodium citrate retards the growth of E.coli.
b) Sodium thiosulfate is used for differentiation of H2S+  and  H2S production.
c) Sodium deoxycholate is a bile salt.
d) Lactose is a source of carbon.
e) Neutral red is a indicator dye.

17 . Triple sugar iron agar
Peptone -0.2 gm
NaCl -0.5 gm
Glucose -0.1 gm
Lactose -1 gm
Sucrose -1 gm
Meat extract -0.3 gm
Yeast extract -0.3 gm
FeSO4 –0.2 gm
Agar-agar -3 gm
Phenol red -0.02 gm
pH -7.4
Sodium thiosulphate -0.03 gm

Role -Triple sugar iron agar is a nutrient rich agar which allows luxurious  growth of all bacteria.

18. Mannitol salt agar
Mannitol -1 gm
Peptone -1 gm
NaCl -0.75 gm
Beef extract -0.1 gm
Phenol red -0.025 gm
Agar-agar -3 gm
Distill water -100 ml

 Role – It is used for isolation of pathogenic strain of Staphylococcus aureus.

19. Egg yolk agar
Peptone -20 gm
Na2HPO4 -2.5 gm
MgSO4 -0.05 gm
NaCl -1 gm
Dextrose -1 gm
Agar – agar -12.5 gm
Distill water -500 ml
pH -7.4
Egg yolk from one egg.

Role – Egg yolk agar is used for isolation of Clostridium and other anaerobic species.

a)  Na2HPO is a buffer.
b) MgSOis a source of Mg++ ion.
c) Dextrose is a fermented sugar it allows luxuriant growth of bacteria.

20. Czapek Dox Thom's agar
Sucrose -3 gm
Sodium nitrate -0.3 gm
K2HPO4 -0.1 gm
Magnesium sulphate -00.5 gm
Potassium chloride -00.5 gm
Ferrous sulphate -Trace amount
Agar-agar -3 gms
Distill water -100 ml
pH -5.4

Role – It is used for isolation and cultivation of fungi.

a) Sucrose is a source of Carbon.
b) Sodium nitrate is a source of nitrogen.
c) K2HPO  acts as a buffer.
d) Ferrous sulphate and acidic pH slow down the growth of bacteria and allows the growth of fungus.
These were few nutrient medium composition for more nutrient medium composition check out my upcoming article Microbial Culture Media Composition Part-3.

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