(ZR) (Z1..

See I)Robertson’s Cooked Meat (RCM) medium is used for the

cultivation of aerobic, microaerophilic, and anaerobic microorganisms,
especially Clostridium species. It is also known as Cooked Meat Broth
(CMB) as it contains pieces of fat free minced cooked meat of ox heart and
nutrient broth. It supports the growth of both spore-forming and non-spore
forming obligate anaerobes and also differentiate between putrefactive and
saccharolytic species.

(Z2) – { Oxygen in culture media can be reduced by various agents such as

glucose, thioglycollate, cooked meat pieces, cysteine and ascorbic acid.

Thioglycollate broth which contains nutrient broth and 1% thioglycollate is

also used to cultivate anerobes. }

(Z3) Principle

Before inoculation RCM/CMB medium is boiled to make it oxygen free. After

inoculation it is covered with a layer of sterile liquid paraffin oil to prevent
entry of oxygen in the medium.

The ingredients present in the medium help to maintain the anaerobic

(reduced) environment :
Unsaturated fatty acids present in meat utilize oxygen for auto-oxidation ;
this reaction is catalysed by haematin in the meat.
Because reducing substances are more available in denatured protein, the
meat particles are cooked before use in the medium.
Glutathione and cysteine (both are reducing agents) present in meat also
utilize oxygen.
Sulphydryl compounds (present in cysteine) also contribute for a reduced
oxidation-reduction potential.
Hemin and vitamin K are added to enhance the growth of anaerobic
microorganisms.
(*for detailed explanation of each component and its significance, read the
composition and functions of ingredients below)

(Z4) Composition
Final pH 6.8 +/- 0.3 at 25ºC.

(Z5) Preparation of the medium

Robertson’s cooked meat medium is best prepared from ready to use
dehydrated granules available from most suppliers of culture media.
Using a small tube or scoop pre-marked to hold 1g of granules, dispense the
medium in 1 g amounts in screw-cap bottles or tubes.
Add 10 ml of distilled water, mix, and allow to soak for 5 minutes.
Sterilize the medium by autoclaving (with caps loosened) at 121°C for 15
minutes. When cool, tighten the bottle caps. Date the medium and give it a
batch number.
Store the medium in a cool dark place, making sure the bottle cops are
tightly screwed.
Shelf-life: 2 years providing there is no change in the volume or appearance
of the medium to suggest contamination.

pH of the medium: This should be within the range pH 7.0-7.4 at room

temperature.

(Z6) Inoculation

Depending on the specimen, cooked meat medium is inoculated using a

swab, Pasteur pipette, or wire loop. If using a swab this should be inserted to
the bottom of the container.

For the culture of strict anaerobes, the medium is best used fresh or after
being placed (with bottle top loosened) in a container of boiling water for
10-15 minutes to drive off any dissolved oxygen or in water bath at 80°C for
30 minutes to make it oxygen free. Allow the medium to cool to room
temperature before inoculating it. The surface of CMB medium may be
covered with a layer of sterile liquid paraffin.

(Z7) Interpretation
A saccharolytic reaction is shown by reddening of the meat with a rancid
smell due to carbohydrate decomposition. A proteolytic reaction is shown by
blacking of the meat with a very unpleasant smells due to protein
decomposition.

Clostridium perfringens: Saccharolytic anaerobes (turn the color of meat

pieces into red)
Clstridium tetani: Proteoltyic anaerobes (blacking of the meat)
(Z8-ZR) Uses of Robertson’s cooked meat medium

Cultivation of aerobic, microaerophilic, and anaerobic microorganisms,

especially Clostridium species. It supports growth of both spore-forming and
non-spore forming obligate anaerobes.
The Food and Drug Administration recommends its use in the enumeration
and identification of Clostridium perfringens from food.