SHEEP BLOOD AGAR BASE, MODIFIED PLATE

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SHEEP BLOOD AGAR​ BASE, MODIFIED PLATE Dehydrated Culture Media

INTENDED USE for cultivation and studying haemolytic reactions of Bacillus cereus in accordance with ISO 21871:2006.

Composition

Ingredients

Gms/Ltr.

Enzymatic digest of casein

15.000

Agar

12.500

Sodium chloride

5.000

Enzymatic digest of soya

5.000

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PRODUCT SUMMARY AND EXPLANATION Sheep Blood Agar​ Base, Modified with added sheep blood was developed to allow maximum recovery of B. cereus without interfering with their haemolytic reactions. This medium is formulated in accordance with ISO. It was formulated to be compatible with sheep blood and give improved haemolytic reactions of organisms.

Bacillus cereus is Gram -positive aerobic or facultatively anaerobic, motile, spore forming, rod shaped bacterium that is widely distributed environmentally. B. cereus is associated mainly with food poisoning it is increasingly reported to be the cause of serious and fatal non- gastrointestinaltrakt.

Hemolysis is very useful for differential characteristics that is best viewed when a bright light is transmitted from behind the plate. Four different types of hemolysis can be described: 1. Alpha-hemolysis (α) – Partial hemolysis that results in a greenish discoloration around the colony.

2. Beta-hemolysis (β) – Complete lysis of red blood cells resulting in a clear zone around the colony.

3. Gamma-hemolysis (γ) – No hemolysis resulting in no change in the medium.

4. Alpha-prime-hemolysis (α′) – A small zone of complete hydrolysis that is surrounded by an area of partial hemolysis.

PRINCIPLE Medium contains nutritional components like enzymatic digest of casein, enzymatic digest of soya and the addition of sodium chloride provides an osmotically balanced medium for bacterial cells. The addition of 5% defibrinated sheep blood allows for the determination of hemolytic reactions, an important differential characteristic.

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INSTRUCTION FOR USE Either streak, inoculate or surface spread the test inoculum aseptically on the plate.

QUALITY CONTROL SPECIFICATIONS Appearance:​ Cherry red coloured opaque gel forms in Petri plates.

Quantity of Medium​: 25ml of medium in 90mm plates. .

INTERPRETATION Cultural characteristics observed after incubation at 30-35 °C for 18-48 hours

Microorganisms

ATCC

Inoculum (CFU/ml)

Bacillus cereus

10876 10^3

Growth

Recovery Rate

Colour of colony

Luxuriant

>=70%

Beta

STORAGE & STABILITY On receipt, store the plates at 2–8 °C. Avoid freezing and overheating. Do not open until ready to use. Prepared plates stored in their original sleeve wrapping until just prior to use may be inoculated up to the expiration date and incubated for recommended incubation times. Allow the medium to warm to room temperature before inoculation.

Product Deterioration:​ Do not use plates if they show evidence of microbial contamination, discoloration, drying, cracking or other signs of deterioration.

REFERENCES 1. MacFaddin JF. Media for isolation-cultivation maintenance of medical bacteria, Vol I. Baltimore: Williams & Wilkins, 1985.

2. Isenberg HD, Ed. Clinical microbiology procedures handbook. Washington, DC: ASM, 1992.

3. Kramer JM, Gilbert RJ. Bacillus cereus and other Bacillus species. In: Doyle MP, ed.

Foodbome Bacterial Pathogens, New York: Marcel Dekker, 1989; 21-70

4 4. NCCLS. Quality assurance of commercially prepared microbiological culture media. 2nd ed. NCCLS document M22-A2. Wayne, PA: NCCLS, 1996.

5. Forbes BA, Sahm DF, Weissfeld AS. Bailey and Scott’s diagnostic microbiology. 10th ed. St. Louis: Mosby, 1998. 5. Pelczar M. J. Jr., Reid R. D., Chan E. C. S., 1977, Microbiology, 4th Ed., Tata McGraw-Hill Publishing Company Ltd, New Delhi.

6. Koneman E. W., Allen S. D., Janda W. M., Schreckenberger P. C., Winn W. C. Jr., 1992, Colour Atlas and Textbook of Diagnostic Microbiology, 4th Ed., J. B. Lippinccott Company.

7. International Organization for Standardization (ISO), Draft ISO 21871:2006 Microbiology of Food & Animal feeding stuffs. Horizontal method for the determination of low numbers of presumptive Bacillus cereus-Most probable number technique and detection method