Supplement with 0.1 ml vitamin K1 1% solution dilluted 1/500 in distilled water and 0.2 ml Na2CO3 10% solution.
PROCEDURE
Regenerate tubes on water at 100 ºC, cool fast then inoculate culture using a pipette and starting from the
bottom of the tube to the surface. Incubate at 37 ºC, up to 2 weeks.
RESULTS
Anaerobes produce turbidity in the medium, mainly in the lower part, aerobes grow close to the surface and
microaerophilics prefer middle part of the tube.
REFERENCES
PROCEDURE
Regenerate tubes on water at 100 ºC, cool fast then inoculate culture using a pipette and starting from the
bottom of the tube to the surface. Incubate at 37 ºC, up to 2 weeks.
RESULTS
Anaerobes produce turbidity in the medium, mainly in the lower part, aerobes grow close to the surface and
microaerophilics prefer middle part of the tube.
REFERENCES
- Handbook Culture Media Merck, 1982.
- Alexandru Rafila: Medii de cultura, medii de transport si conservare. In Dumitru Buiuc, Marian Negut:
Tratat de microbiologie clinica ed. III, 2009, 1045-1084.
COMPOSITION
VARIANT 1
Proteose peptone …………………….............………………….. 10.0 g
Meat infusion …………………………................………………..... 5.0 g
D(+) glucose …………………………...............……………...…… 5.0 g
K2HPO4 ………………....................………………………………… 2.0 g
Sodium thioglycollate ……………………................……………… 0.5 g
Methylene blue ……………………………..............…………... 0.002 g
Agar - agar ……………………………………...............…………. 0.5 g
Distilled water ad 1000 ml, pH 7.0 ± 0.1, Pour in tubes and autoclave.
Clear, colorless & semi-solid state. Store in the dark, at room temperature.
Do not use green-blue tubes if color do not disappear after boiling.
VARIANT 2 - for fastidious anaerobes
Casein pancreatic digest …………….............………………….. 17.0 g
Soy pancreatic digest …………….............…………….......…….. 3.0 g
NaCl ….................……………………................………………..... 2.5 g
Glucose ……………………........……...............……………...…... 6.0 g
Sodium sulfite ………....................………………………………… 0.1 g
Sodium thioglycollate ……………………................……………… 0.5 g
Methylene blue ….…………………………..............…………... 0.002 g
Agar ...........……………………………………...............…………. 0.7 g
L-cystine .................................................................................. 0.25 g
Hemin ..................................................................................... 0.005 g
Distilled water ad 1000 ml, pH 7.2, Pour in tubes. Do not boil, do not
autoclave.
VARIANT 1
Proteose peptone …………………….............………………….. 10.0 g
Meat infusion …………………………................………………..... 5.0 g
D(+) glucose …………………………...............……………...…… 5.0 g
K2HPO4 ………………....................………………………………… 2.0 g
Sodium thioglycollate ……………………................……………… 0.5 g
Methylene blue ……………………………..............…………... 0.002 g
Agar - agar ……………………………………...............…………. 0.5 g
Distilled water ad 1000 ml, pH 7.0 ± 0.1, Pour in tubes and autoclave.
Clear, colorless & semi-solid state. Store in the dark, at room temperature.
Do not use green-blue tubes if color do not disappear after boiling.
VARIANT 2 - for fastidious anaerobes
Casein pancreatic digest …………….............………………….. 17.0 g
Soy pancreatic digest …………….............…………….......…….. 3.0 g
NaCl ….................……………………................………………..... 2.5 g
Glucose ……………………........……...............……………...…... 6.0 g
Sodium sulfite ………....................………………………………… 0.1 g
Sodium thioglycollate ……………………................……………… 0.5 g
Methylene blue ….…………………………..............…………... 0.002 g
Agar ...........……………………………………...............…………. 0.7 g
L-cystine .................................................................................. 0.25 g
Hemin ..................................................................................... 0.005 g
Distilled water ad 1000 ml, pH 7.2, Pour in tubes. Do not boil, do not
autoclave.
DESCRIPTION
Medium for the cultivation of anerobic, microaerophilic & facultatively anaerobic germs. Thioglycollate ensures the
anaerobiosis. Increase in oxygen content is indicated by the methylene blue which changes color to green-blue.
Medium for the cultivation of anerobic, microaerophilic & facultatively anaerobic germs. Thioglycollate ensures the
anaerobiosis. Increase in oxygen content is indicated by the methylene blue which changes color to green-blue.
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