Bacteriological Examinations of Stool
Bacteriological Examinations of Stool
Bacteriological Examinations of Stool
Fecal samples should be collected in the early stages of the disease before
antibiotic treatment is instituted, when the pathogenic germs are likely to be
present in large numbers in stools. We will collect the samples stools in a sterile
container of appropriate size, fitted with a tight-fitting cap and waterproof.
Upon receipt at the laboratory, the stools are processed as quickly as possible,
in all case no more than 2 hours after collection.
Principle:
This medium is used for the isolation of salmonella and shigella from stools and
foodstuffs. The development of secondary flora is slowed by brilliant green,
bile salts, the high concentration of thiosulfate and citrate. Lactose, a reactive
sugar in the environment/medium, allows detection of possible growth of
coliforms (red colonies). In addition, bacteria capable of producing H2S by
reduction of thiosulfate give in the presence of iron ions colonies with a black
center.
Medium:
Put 62g of dry medium in a liter of distilled water. Leave to rest for 5 minutes,
then shake to obtain a homogeneous suspension. Heat gently, stirring
occasionally for 1 to 2 minutes. Do not autoclave. Leave to cool to around 45°C
and pour into a petri dish, of approximately 20ml per box.
Allow the medium to solidify, with the can lid slightly raised.
It should be noted that any modification in the concentration of the medium
(prolonged boiling for dissolution, significant drying of the boxes etc.) makes
the environment very inhibitory, hence an often delicate preparation,
disappointing results and the preference generally given to ready-for-use
medium.
Reading / Results:
Colonies- lactose-positive red colonies.
Lactose-negative colonies: colorless colonies
H2S-positive colonies: colonies with a black center.
The differential identification of Shigella will be considered from colorless
colonies, that of salmonella from colorless colonies with or without a black
center.
II.3.2.MacConkey Medium: 11.4.2
Principle:
The selenite present in the medium inhibits the growth of coliform bacteria and
enterococci within 12 hours after the start of incubation; then the inhibitory
action slowly decreases. Salmonella and Proteus, on the other hand, are little
inhibited from the start. It is therefore not recommended to extend the
incubation time.
Medium:
A.IPP Formula
Dissolve 4g of sodium selenite in a liter of distilled water, then add 19g of the
medium following basis:
Bacteriological peptone...5g
Disodium phosphate...10g
Lactose…4g
PH: 7
Heat to dissolve. Mix well, distribute at the rate of 10ml per tube (screw cap).
Sterilize in a boiling water bath or in flowing steam for 30 minutes. Don’t
autoclave. It is also possible to sterilize by membrane filtration. Distribute into
sterile tubes.
A. Formula (Merck, BioMérieux, Oxoid, Difco)
Selenite is incorporated into the basic medium whose formula (in grams per
liter) is
Special Peptone…5.0g
Lactose…4.0g
Sodium selenite… 4.0g
Dipotassium phosphate... 3.5g
Monopotassium phosphate...6.5g