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Bile Esculin HiVeg™ Agar

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MV972

Recommended for isolation and presumptive identification of group D Streptococci from food and pharmaceutical product.

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Description

Intended Use

Bile Esculin HiVeg Agar / Agar Base is a differential medium recommended for isolation and presumptive identification of Group D Streptococci from food and pharmaceutical products.

Composition

Ingredients	MV972 Grams/Litre	MV340 Grams/Litre
HiVeg peptone	25.00	22.00
HiVeg extract	6.00	6.00
Synthetic detergent No.Il	2.00	5.00
HiVeg hydrolysate	15.00	15.00
Esculin	1.00	-
Ferric citrate	0.50	0.50
Agar	15.00	15.00
Final pH (at 25°C)		6.6 ± 0.2

** Formula adjusted, standardized to suit performance parameters

Product Profile

Vegetable based (Code MV)	Animal based (Code M)
MV972/MV340	M972/M340
HiVeg peptone	Peptic digest of animal tissue
HiVeg extract	Beef extract
Synthetic detergent No.Il	Oxgall
HiVeg hydrolysate	Casein enzymic hydrolysate

Reconstitution

(MV972) : 64.5 g/l
(MV340) : 63.5 g/l

Quantity on preparation (500g)

(MV972) : 7.75 L
(MV340) : 7.87 L

(100g)

(MV972) : 1.55 L

pH (25°C)

6.6 ± 0.2

Supplement

(MV340) : Esculin (FD050)

Sterilization

121°C / 15 minutes.

Storage

Dry Medium Below 30°C, Prepared Medium 2-8°C.

Directions

Suspend 64.5 grams of MV972 or 63.5gms of MV340 in 1000 ml distilled water. Heat to boiling to dissolve the medium completely. Add 1 gram of esculin (2 vials of FD050) in MV340. Mix and dispense into tubes or flasks as desired. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Allow the tubed medium to solidify in slanted position.

Principle and Interpretation

This medium is prepared by completely replacing animal based peptones with vegetable peptones which are free from BSE/TSE risks. Bile Esculin HiVeg Agar is the modification of Bile Esculin Agar which was formulated by Swan (1) for the isolation and identification of Group D Streptococci from food. The medium contains Synthetic detergent No. II that inhibits gram positive bacteria other than group D Streptococci and Enterococci. Enterococci and Group D Streptococci were able to split esculin to esculetin and dextrose, which reacts with ferric citrate producing brownish black precipitate (2). Ferric citrate is incorporated into the medium as an indicator of esculin hydrolysis and resulting esculatin formation. Originally Bile Esculin Test was used for identification of Enterococci, but it was found that this test is also shared by Group D Streptococci (3) and therefore it is recommended that other tests such as salt tolerance be performed while identifying Enterococci (4). Similarly this medium was also shown to aid differentiation of Enterobacteriaceae, Klebsiella, Enterobacter-Serratia division from other Enterobacteriaceae genera (5) on the basis of esculin hydrolysis. Occasional strains of viridans Streptococci blacken the medium or display weakly positive reactions (6).

Quality Control

Appearance of Powder
Brownish yellow coloured may have slightly greenish tinge, homogeneous, free flowing powder.

Gelling
Firm, comparable with 1.5% Agar gel.

Colour and Clarity
Yellow coloured, clear to slightly opalescent gel with a bluish tinge forms in petri plates.

Reaction
Reaction of 6.45% w/v of MV972 or 6.35% w/v of MV340 aqueous solution is pH 6.6 ± 0.2 at 25°C.

Cultural Response
Cultural characteristics observed after an incubation at 35 - 37°C for 18-24 hours in an increased atmosphere of carbon dioxide

Organisms (ATCC)	Inoculum (CFU)	Growth	Recovery	Esculin hydrolysis
Enterococcus faecalis (29212)	10²-10³	luxuriant	>50%	+
Sreptococcus pyogenes (19615)	10²-10³	none-poor	<10%	-
Proteus mirabilis (25933)	10²-10³	luxuriant	>50%	-

Key: + = Blackening of the medium
- = No Change

Product Information

More Information
Product Name	Bile Esculin HiVeg™ Agar
SKU	MV972
Product Type	HiVeg™
Physical Form	Powder
Origin	Animal Free (Veg)
Packaging type	HDPE
References	1. Koneman E. W., Allen S. D., Janda W. M., Schreckenberger P. C., Winn W. C. Jr., 1992, Colour Atlas and Textbook ofDiagnostic Microbiology, 4 th Ed., J. B. Lippinccott Company2.Meyer and Schonfeld, 1926, Zentralbl. Bakeriol, Parasitenk. Infectionskr. Hyg. Abt. Orig. 99:402.3.MacFaddin J. F., 1985, Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria, Vol. I, Williamsand Wilkins, Baltimore.4.Rochaix, 1924, Comt. Rend. Soc. Biol., 90:771.5.Facklam R., 1973, Appl. Microbiol., 26:138.Swan, 1954, J. Clin. Pathol., 7:160.6.Facklam R., 1972, Appl. Microbiol., 23:1131.7.Facklam R. R and Moody M. D., 1970, Appl. Microbiol., 20(2):245.8.Edberg S. C., Pittman S., and Singer J. M., 1977, J. Clin. Microbiol., 6:111.9.Murray P. R., Baron E. J., Jorgensen J. H., Pfaller M. A., Yolken R. H.,(Eds.), 8th Ed., 2003, Manual of Clinical Microbiology,ASM, 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.American Public Health Association, Standard Methods for the Examination of Dairy Products, 1978, 14th Ed., WashingtonD.C.13.Salfinger Y., and Tortorello M.L. Fifth (Ed.), 2001, Compendium of Methods for the Microbiological Examination ofFoods, 5th Ed., American Public Health Association, Washington, D.C.14.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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