CHOCOLATE BACITRACIN (HAEMOPHILUS SELECTIVE) AGAR

Technical Data #1984a / 98.06.16

Chocolate Bacitracin Agar is a selective enriched medium for the isolation of Haemophilus sp. Hemoglobin and Bio-X enrichment replace lysed heated blood and give to the culture media enough nutrients agents to permit luxuriant growth of Haemophilus Bacitracin reacts as a selective agent and prevents normal flora growth.

FORMULA
in grams per liter purified filtered water

Casein Peptone

7,5

Meat Peptone

7,5

Corn Starch

1,0

Potassium Phosphate, dibasic

4,0

Potassium Phosphate, monobasic

1,0

Sodium Chloride

5,0

Agar

10,0

Hemoglobin

10,0

Enrichment Bio-X

10 ml

Antibiotic  
Bacitracin

0,3

pH 7,2 +/- 0,2 à 25º C

This approximate formula may be adjusted and/or enriched to obtain best results.


PRECAUTIONS
This medium is for IN VITRO diagnostic use only.


STORAGE
Store prepared media at 2-8º C protected from direct light and dehydrated powder, in a dry place, in tightly-sealed containers at 2-25º C.


SIGN OF DETERIORATION
Media should not be used if the expiry date has passed. Prepared media should not be used if there are signs of contamination or deterioration (shrinking, cracking, evaporation or discoloration). Do not use dehydrated media if it is caked.


DIRECTIONS
Suspend 36 g of dehydrated media in 500 ml of purified filtered water. Heat with frequent agitation and boil for one minute. Sterilize at 121º C for 15 minutes. Cool to 45-50º C. Add 500 ml of sterile 2% Hemoglobin (8662), 10 ml of Bio-X Enrichment (8601) and 0,3 g of Bacitracin solution. Mix gently and dispense into sterile Petri dishes.


PROCEDURE
Prior to inoculate, the prepared media should be brought to room temperature.

  1. Directly, inoculate plate as soon as possible with specimen and streak for isolation.
  2. Incubate at 35º C with 5% CO2 for 24 hours.


QUALITY CONTROL
Results after 24 hrs at 35ºC W CO2

Organisms

ATCC

Growth

Haemophilus influenzae

10211

+

Escherichia coli

25922

- or partial

Staphylococcus aureus

25923

-

On Chocolate Bacitracin Agar, Haemophilus influenzae colonies are small, colorless, moist and transparent.


LIMITATIONS OF METHOD
This medium is only a part of the identification. Other tests may be required.


REFERENCES

  1. Chapin, K.C., G.V. Doern 1983 Selective media for recovery of Haemophilus influenzae from specimens contaminated with upper respiratory tract microbial flora. J. Clin. Microbiol. 17:1163-1165
  2. Lennette, E.H., Ballows, A., Hausler, W.J.Jr., and Shadomy, H.J. Manual of Clinical Microbiology. 4th ed. 1985 Washington D.C.: American society for Microbiology.


CATALOGUE NUMBERS

Dehydrated media 500 g QB-39-1906
Hemoglobin 500 g QB-39-2114
     
Prepared media    
Plates, 100x15 mm 10/pkg 1984
Mono-Plates, 100x15 mm 10/pkg 1984M
     
Bio-X Enrichment, 10 ml 10/pkg 8601
     
Hemoglobin, 2% sterile 500 ml 8662

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