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Bile Esculin HiVeg™ Agar Base

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MV340
For isolation and presumptive identification of group D Streptococci from food and pharmaceutical products.

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.

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

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

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).

Technical Details

Reconstitution:

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

Quantity on preparation:

  • (500g): (MV972): 7.75 L
  • (500g): (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.

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

References

  1. Swan A., 1954, J. Clin. Pathol., 7:160.
  2. MacFaddin J.F., 1985, Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria, Vol. 1, Williams and Wilkins, Baltimore.
  3. Facklam R., 1972, Appl. Microbiol., 23:1131.
  4. Facklam R., 1973, Appl. Microbiol., 26:138.
  5. Edberg S.C., Pittman S., and Singer J.M., 1977, J. Clin. Microbiol., 6:111.
  6. Facklam, etal 1999. In Murray, Baron, pfaller, Tenover and yolken (ed.), Manual of clinical Microbiology, 7th ed. ASM, Washington, D. C.
More Information
Product Name Bile Esculin HiVeg™ Agar Base
SKU MV340
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 Company
2.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.
6.Swan, 1954, J. Clin. Pathol., 7:160.
7.Facklam R., 1972, Appl. Microbiol., 23:1131.
8.Facklam R. R and Moody M. D., 1970, Appl. Microbiol., 20(2):245.
9.Edberg S. C., Pittman S., and Singer J. M., 1977, J. Clin. Microbiol., 6:111.
10.Murray P. R., Baron E. J., Jorgensen J. H., Pfaller M. A., Yolken R. H.,(Eds.), 8th Ed., 2003, Manual of ClinicalMicrobiology, ASM, Washington, D.C.
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