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Microbial Culture Media : Pharmaguideline https://www.pharmaguideline.com/2011/02/culture-media.html

Microbial Culture Media

Know the composition of all microbiological media used for microbiological analysis.
Baird Parker Agar Base
Specification
Solid and selective culture medium for the screening of staphylococci from a variety of samples, acc. to pharmacopeias and ISO standard.
Formula (in g/L)
Tryptone ...................................................10,0
Sodium pyruvate ......................................10,0
Glycine .....................................................12,0
Meat extract ................................................5,0
Lithium chloride ..........................................5,0
Yeast extract ...............................................1,0
Agar .........................................................17,0
Final pH 7,0 ± 0,2

Directions
Suspend 60 g in 950 mL of distilled water. Allow it to soak and then bring to the boil with constant stirring. Sterilize by autoclaving at 121° C for 15
minutes. Cool to 50° C and add 50 mL of Egg’s Yolk Tellurite Sterile Emulsion (Ref. 06-026). Homogenize and distribute into plates. Once prepared,
the medium must not be reheated nor sterilized again.
Description
The Baird Parker Agar Base is specially recommended for the detection and enumeration of staphylococci in food and other material, since it allows
a good differentiation of coagulase-positive strains. The growth of the accompanying bacteria is usually suppressed by the high concentration in
lithium, glycine and pyruvate. Lithium and glycine enhances the growth of staphylococci. Even if its high selectivity does not affect staphylococci it
may occasionally permit the growth of some Bacillus species, yeast and very rarely, Proteus. The growth of Proteus species can be suppressed by
adding 50 mg/l of sulphamethazine.
The presence of tellurite and egg’s yolk, which must always be added to the medium after sterilization, allows the differentiation of presumptly
pathogenic staphylococcal colonies. A high correlation has been found between the coagulase test and the presence of clearing zones of lypolysis in
this medium, which is due to the staphylococcal lecithinase. On the other hand, studies show that almost 100% of coagulase-positive staphylococci
are capable of reducing tellurite, which produces black colonies, whereas other staphylococci can not always do so.
When using sterile reagents other than SCHARLAU MICROBIOLOGY, the complete medium may be obtained by adding 50 mL sterile egg’s yolk
and 10 mL of 1% potassium tellurite solution. Plates should be used on the same day of preparation or within 48 hours, to avoid the loss of definition
in the precipitated zones . The basal medium, without the yolk or the tellurite, is perfectly stable and therefore can be repeatedly melted.
Technique
The inoculation is done by spreading 0,5 mL of sample over each plate with a Drigalski loop (Ref. 5-010). After 18-24 hours of incubation at 35° C,
select the colonies which are black, shiny and convex with regular margins surrounded by a zone of clearing. These can be presumptly identified as
coagulase-positive Staphylococcus aureus.
Colonial appearance after 24 h. at 35°C:
Staphylococcus aureus: Black, shiny, convex, regular margins, 1.0-1.5 mm diameter, surrounded by a clearing zone of lipolysis 2-5 mm in width. May
produce wide opaque precipitate zones extending into the cleared medium after 48 hours.
Other species of Staphylococcus: Black, usually dully, with regular margins. Sometimes they are brown with zones of clearing but these present wide
opaque zones.
Micrococcus spp: Brown, very small and without clearing.
Bacillus spp: Various shades of brown, big. May produce clearing after 48 hours.
Yeasts: White, big and smooth.
The egg’s yolk emulsion can be prepared by mixing a fresh egg’s yolk with an equivalent quantity (w/v) of saline solution. Sterilize by filtration and
aseptically add to the medium. This reagents´s reference, already sterilized, is SCHARLAU 6-016.
The potassium tellurite solution is prepared by dissolving 3,5 g potassium tellurite in 100 mL distilled water. Sterilize by filtration. This sterile reagent’s
SCHARLAU MICROBIOLOGY reference is 6-011.
Although these solutions can be mixed to be added to the Baird Parker Agar Base forming the commonly known Egg Yolk Tellurite Sterile Emulsion
(Ref. 06-026), they are also stable as the separate supplement and can be used in many other culture media.

Blood Agar Base (Columbia)

Specification
Medium especially rich in peptone, appropriate for blood addition or to prepare Chocolate Agar.
Formula (in g/L)
Casein Peptone ........................................12,0
Meat peptone ...........................................11,0
Starch .........................................................1,5
Sodium Chloride .........................................5,0
Agar ..........................................................15,0
Final pH 7,3 ± 0,2
Directions

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