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Bacillus Cereus HiVeg™ Agar Base

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MV833

Used as a selective medium for isolation, detection and enumeration of Bacillus cereus.

Description

Intended Use

Bacillus Cereus HiVeg Agar Base with added supplements is used as a selective medium for the isolation and enumeration of Bacillus cereus.

Composition

Ingredients	Grams/Litre
HiVeg peptone	1.0
Mannitol	10.0
Sodium chloride	2.0
Magnesium sulphate	0.1
Disodium phosphate	2.5
Monopotassium phosphate	0.25
Sodium pyruvate	10.0
Bromo thymol blue	0.12
Agar	15.0
Final pH (at 25°C)	7.2 ± 0.2

** Formula adjusted, standardized to suit performance parameters.

Product Profile

Vegetable based (Code MV)	MV833 HiVeg peptone
Animal based (Code M)	M833 Peptic digest of animal tissue
Recommended for	Selective medium for the isolation and enumeration of Bacillus cereus.
Reconstitution	41.0 g/l
Quantity on preparation (500g)	12.19 L
Quantity on preparation (100g)	2.43 L
pH (25°C)	7.2 ± 0.2
Supplement	Polymyxin B Selective Supplement (FD003) Egg Yolk Emulsion (FD045)
Sterilization	121°C / 15 minutes.
Storage	Dry Medium - Below 30°C, Prepared Medium 2 - 8°C.

Directions

Suspend 20.5 grams in 475 ml distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 50°C and aseptically add rehydrated contents of 1 vial of Polymyxin B Selective Supplement (FD003) and 25 ml of sterile Egg Yolk Emulsion (FD045). Mix well and pour into sterile petri plates.

Principle and Interpretation

Bacillus Cereus HiVeg Agar is developed by using HiVeg peptone, which is free of BSE/TSE risks. This medium is the modification of Bacillus Cereus Agar Base developed by Holbrook and Anderson (1), which is a highly specific, selective medium for the isolation and enumeration of Bacillus cereus from foods. It supports the growth of even a small number of Bacillus cereus cells and spores in the presence of large number of other food contaminants. The typical colonies of Bacillus cereus are crenated, about 5 mm in diameter and have a distinctive turquoise to peacock blue colour surrounded by a good egg yolk precipitate of the same colour.

Addition of Polymyxin B Sulphate (2, 3) at a final concentration of 100 units per ml of medium is sufficient to make the medium selective for the isolation of Bacillus cereus. If moulds are suspected in the inoculum, 40 mcg per ml of filter-sterilized, Cycloheximide may be incorporated to suppress the mould contamination. Some strains of Bacillus cereus have very weak egg yolk reaction. Moreover, on this medium Bacillus cereus is indistinguishable from Bacillus thuringiensis. Bacillus cereus causes food poisoning due to the consumption of contaminated rice (4, 5, 6), eye infections and a wide range of other clinical conditions like abscess formation, meningitis, septicaemia and wound infection. Bacillus cereus is a known cause of disease mastitis, especially in ewes and heifers among the veterinarians. HiVeg peptone and sodium pyruvate improve egg yolk precipitation and enhance sporulation. Bromo thymol blue act as pH indicator to detect mannitol fermentation.

Quality Control

Appearance of powder
Greenish yellow coloured, homogeneous, free flowing powder.

Gelling
Firm, comparable with 1.5% Agar gel.

Colour and Clarity
Basal medium yields green coloured clear to slightly opalescent gel. With additon of 5% egg yolk emulsion yellowish green coloured opaque gel forms in petri plates.

Reaction
Reaction of 4.1% w/v aqueous solution of basal medium is pH 7.2 ± 0.2 at 25°C.

Cultural Response
Cultural characteristics observed after an incubation at 35 - 37°C for 24 hours with added Polymyxin B Selective Supplement (FD003) and Egg Yolk Emulsion (FD045).

Organisms (ATCC)	Inoculum (CFU)	Growth	Recovery	Colour of colony	Egg Yolk Reaction
Bacillus cereus (10876)	102-103	good-luxuriant	>50%	blue	precipitation
Escherichia coli (25922)	102-103	inhibited	0%	-	-
Proteus vulgaris (13315)	102-103	good-luxuriant	>50%	green	clearing
Serratia marcescens (8100)	102-103	good-luxuriant	>50%	red	clearing
Staphylococcus aureus (25923)	102-103	good-luxuriant	>50%	yellow	clearing

References

Holbrook R. and Anderson J., 1980, Can. J. Microbiol., 26(7):753.
Donovan K.O., 1958, J. Appl. Bacteriol, 21(1):100.
Mossel, D.A.A., Koopman J. and Jongerius E., 1967, J. Appl. Microbiol., 15(3):650.
Mortimer P.R. and McCann. G, 1974, Lancet, 1043.
Bouza E., Grant S., Jordan C., et al, 1979, Arch. Ophthalmol., 97:488.
Wohlgemuth K., Kirkbride, C.A., Bicknell, E. J. and Ellis, R.P., 1972, Am. Vet. Med. Ass., 161:1691.

Product Information

More Information
Product Name	Bacillus Cereus HiVeg™ Agar Base
SKU	MV833
Product Type	HiVeg™
Physical Form	Powder
Origin	Animal Free (Veg)
Packaging type	HDPE
References	1. Holbrook R. and Anderson J., 1980, Can. J. Microbiol. 26(7):753-7592.Donovan K.O., 1958, J. Appl. Bacteriol, 21(1): 100.3.Mossel D.A.A., Koopman J. and Jongerius E., 1967, J. Appl. Microbiol. 15(3):650-653.4.Mortimer P.R. and McCann G., 1974, Lancet, 1043-1045.5.Bouza E., Grant S., Jordan C. et al, 1979, Arch. Ophthalmol. 97:4986.Wohlgemuth K., Kirkbride, C.A., Bicknell, E. J. and Ellis, R.P., 1972 , J. Am. Vet. Med. Ass. 161:1691-1695.7.Kirnbull P.C., J. Clin. Pathol. 32:2898.American Public Health Association, Standard Methods for the Examination of Dairy Products, 1978, 14th Ed., WashingtonD.C.9.Salfinger Y., and Tortorello M.L. Fifth (Ed.), 2015, Compendium of Methods for the Microbiological Examination ofFoods, 5th Ed., American Public Health Association, Washington, D.C.10.Isenberg, H.D. Clinical Microbiology Procedures Handb0ook. 2nd Edition.11. Jorgensen,J.H., Pfaller , M.A., Carroll, K.C., Funke, G., Landry, M.L., Richter, S.S and Warnock., D.W. (2015)Manual of Clinical Microbiology, 11th Edition. Vol. 1.12.Wehr H. M. and Frank J. H., 2004, Standard Methods for the Microbiological Examination of Dairy Products, 17th Ed.,APHA Inc., Washington, D.C.
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