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TITAN MEDIA

ANAEROBIC AGAR                                                                             TM 010
For cultivation of anaerobic bacteria especially Clostridium species

Composition

                    Ingredients                          Gms/Ltr.
           Casein enzymatic hydrolysate                     20.00
                       Agar                                 20.00
                     Dextrose                               10.00
                 Sodium chloride                            5.00
               Sodium thioglycollate                        2.00
        Sodium formaldehyde sulphoxylate                    1.00
                  Methylene blue                            0.002

* Dehydrated powder, hygroscopic in nature, store in a dry place, in tightly-sealed containers below 25°C
and protect from direct Sunlight.

Instructions for Use
Dissolve 58gms in 1000ml of distilled water. Gently heat to boiling with gentle swirling and dissolve the
medium completely. Sterilize by autoclaving at 15 psi (121°C) for 15 minutes. Mix well. Cool to 45-500C
and pour into sterile Petri plates.

Appearance: Light amber colour, clear to slightly opalescent gel
pH (at 25°C): 7.2 ± 0.2

Principle
ANAEROBIC AGAR is used for cultivation of anaerobic bacteria especially Clostridium species. Medium
contains Casein enzymatic hydrolysate as a source of nitrogen, vitamins & minerals. Dextrose is a carbon
energy source. Sodium thiglycollate and Sodium formaldehyde sulphoxylate provides a low oxidationreduction
potential, and securing anaerobic conditions as indicated by Methylene blue present in the
medium which yields blue colour to medium in presence of oxygen. Sodium chloride helps to maintain
the osmotic balance of the medium. Agar is the solidifying agent. For testing standard petri plates or
Anaerobic Agar, Brewer’s (TM 011) plates with the medium dispense 50-60ml medium and cool at 45-
500C by surface inoculation or by pour plating technique. Sometimes, on slightly heat fixing the sample
when spore formers Clostridium are present, except C. perfringens (which rarely forms spores).

Interpretation
Cultural characteristics observed under anaerobic conditions after inoculating (103CFU/ml), on
incubation at 35 ± 2°C for 48 - 72 hours.


      Microorganisms                       ATCC                         Inoculum                        Growth
                                                                         (CFU/ml)

   Clostridium perfringens                  12924                          10³                              Good
   Clostridium sporogenes                   11437                          10³                              Good
    Clostridium butyricum                   13732                          10³                              Good
References
1.Brewer J. H., Science, 95:587. (1942).
2. Baron E. J., Peterson and Finegold S. M., Bailey & Scotts Diagnostic Microbiology, 9th Ed.,
Mosby-Year Book, Inc., St. Louis, Mo. (1994).

For more details visit:- http://www.titanmedia.in/images/upload/td/TM%20010_ANAEROBIC%20AGAR%20TM
%20010.pdf or http://www.titanmedia.in

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ANAEROBIC AGAR

  • 1. TITAN MEDIA ANAEROBIC AGAR TM 010 For cultivation of anaerobic bacteria especially Clostridium species Composition Ingredients Gms/Ltr. Casein enzymatic hydrolysate 20.00 Agar 20.00 Dextrose 10.00 Sodium chloride 5.00 Sodium thioglycollate 2.00 Sodium formaldehyde sulphoxylate 1.00 Methylene blue 0.002 * Dehydrated powder, hygroscopic in nature, store in a dry place, in tightly-sealed containers below 25°C and protect from direct Sunlight. Instructions for Use Dissolve 58gms in 1000ml of distilled water. Gently heat to boiling with gentle swirling and dissolve the medium completely. Sterilize by autoclaving at 15 psi (121°C) for 15 minutes. Mix well. Cool to 45-500C and pour into sterile Petri plates. Appearance: Light amber colour, clear to slightly opalescent gel pH (at 25°C): 7.2 ± 0.2 Principle ANAEROBIC AGAR is used for cultivation of anaerobic bacteria especially Clostridium species. Medium contains Casein enzymatic hydrolysate as a source of nitrogen, vitamins & minerals. Dextrose is a carbon energy source. Sodium thiglycollate and Sodium formaldehyde sulphoxylate provides a low oxidationreduction potential, and securing anaerobic conditions as indicated by Methylene blue present in the medium which yields blue colour to medium in presence of oxygen. Sodium chloride helps to maintain the osmotic balance of the medium. Agar is the solidifying agent. For testing standard petri plates or Anaerobic Agar, Brewer’s (TM 011) plates with the medium dispense 50-60ml medium and cool at 45- 500C by surface inoculation or by pour plating technique. Sometimes, on slightly heat fixing the sample when spore formers Clostridium are present, except C. perfringens (which rarely forms spores). Interpretation Cultural characteristics observed under anaerobic conditions after inoculating (103CFU/ml), on incubation at 35 ± 2°C for 48 - 72 hours. Microorganisms ATCC Inoculum Growth (CFU/ml) Clostridium perfringens 12924 10³ Good Clostridium sporogenes 11437 10³ Good Clostridium butyricum 13732 10³ Good
  • 2. References 1.Brewer J. H., Science, 95:587. (1942). 2. Baron E. J., Peterson and Finegold S. M., Bailey & Scotts Diagnostic Microbiology, 9th Ed., Mosby-Year Book, Inc., St. Louis, Mo. (1994). For more details visit:- http://www.titanmedia.in/images/upload/td/TM%20010_ANAEROBIC%20AGAR%20TM %20010.pdf or http://www.titanmedia.in