HI Agar, HiVeg® (Heart Infusion Agar, HiVeg®)

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MV169
Recommended for the isolation and cultivation of a wide variety of fastidious organisms. It is also used as a base for preparation of blood agar in determining haemolytic reactions.


Intended Use

Recommended for the isolation and cultivation of a wide variety of fastidious organisms. It is also used as a base for preparation of blood agar in determining haemolytic reactions.

Composition**

Ingredients g / L
HiVeg® infusion 10.000
HiVeg® hydrolysate No. 1 10.000
Sodium chloride 5.000
Agar 15.000
Final pH (at 25°C) 7.4±0.2

**Formula adjusted, standardized to suit performance parameters

Directions

Suspend 40.0 grams in 1000 ml of purified / distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 45-50°C. If desired 5% v/v sterile defibrinated blood may be added. Mix well and pour into sterile Petri plates.

Principle And Interpretation

HI Agar, HiVeg® is prepared by using vegetable peptones in place of animal based peptones which make the media free of BSE/TSE risks. Fastidious organisms having exacting nutritional requirement could be cultivated on infusion media, as demonstrated by Huntoon (1). A liquid medium containing an infusion of meat was one of the first media used for the cultivation of bacteria. These infusion media need not be further supplemented by the addition of supplements for cultivation of fastidious bacteria (2). HI Agar, HiVeg®, containing HiVeg® infusion from (equivalent to infusion from beef heart) is used for the isolation and cultivation of a wide variety of fastidious organisms (3). HI Agar, HiVeg® can also be used for the cultivation of Vibrio species (2,4). It can also be supplemented with glucose, horse serum and antibiotics for the cultivation a wide variety of organisms (3). It is used for mass cultivation of organisms for preparation of vaccines. On supplementation of blood, HI Agar, HiVeg® can be used to study haemolytic reactions (6). This medium was used for isolation and enumeration of haemolytic Streptococci in milk (5). HiVeg® infusion and HiVeg® hydrolysate No. 1 infusion provide nutritional requirements for the pathogenic bacteria. Sodium chloride maintains the osmotic equilibrium of the medium.

Type of specimen

Food and dairy samples - Milk, meat samples

Specimen Collection and Handling

For food samples, follow appropriate techniques for sample collection and processing as per guidelines (4,7). After use, contaminated materials must be sterilized by autoclaving before discarding.

Warning and Precautions

Read the label before opening the container. Wear protective gloves/protective clothing/eye protection/ face protection. Follow good microbiological lab practices while handling specimens and culture. Standard precautions as per established guidelines should be followed while handling specimens. Safety guidelines may be referred in individual safety data sheets.

Limitations :

  1. Further biochemical and serological tests must be carried out for further identification.

Performance and Evaluation

Performance of the medium is expected when used as per the direction on the label within the expiry period when stored at recommended temperature.

Quality Control

Appearance
Cream to yellow homogeneous free flowing powder

Gelling
Firm, comparable with 1.5% Agar gel

Colour and Clarity of prepared medium
Basal medium: Light yellow coloured, clear to slightly opalescent gel After addition of 5-7% w/v sterile defibrinated blood : Cherry red coloured, opaque gel forms in Petri plates

Reaction
Reaction of 4.0% w/v aqueous solution at 25°C. pH : 7.4±0.2

pH
7.20-7.60

Cultural Response
Cultural characteristics observed with added 5-7% w/v sterile defibrinated blood, after an incubation at 35-37°C for 18-48 hours.

Organism Inoculum (CFU) Growth w/o blood Recovery w/o blood Growth with blood Recovery with blood Haemolysis
Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) 50-100 good-luxuriant >=70% luxuriant >=70% beta
Neisseria meningitidis ATCC 13090 50-100 luxuriant >=70% luxuriant >=70% none
Streptococcus pneumoniae ATCC 6303 50-100 good 50-70% luxuriant >=70% alpha
Streptococcus pyogenes ATCC 19615 50-100 good 50-70% luxuriant >=70% beta
Escherichia coli ATCC 25922 (00013*) 50-100 luxuriant >=70% luxuriant >=70% beta

Key: *Corresponding WDCM numbers.

Storage and Shelf Life

Store between 10-30°C in a tightly closed container and the prepared medium at 2-8°C. Use before expiry date on the label. On opening, product should be properly stored dry, after tightly capping the bottle in order to prevent lump formation due to the hygroscopic nature of the product. Improper storage of the product may lead to lump formation. Store in dry ventilated area protected from extremes of temperature and sources of ignition. Seal the container tightly after use. Product performance is best if used within stated expiry period.

Disposal

User must ensure safe disposal by autoclaving and/or incineration of used or unusable preparations of this product. Follow established laboratory procedures in disposing of infectious materials and material that comes into contact with sample must be decontaminated and disposed of in accordance with current laboratory techniques (8,9).

More Information
Product Name HI Agar, HiVeg® (Heart Infusion Agar, HiVeg®)
SKU MV169
Product Type HiVeg™
Physical Form Powder
Origin Animal Free (Veg)
Packaging type HDPE
References 1. American Public Health Association, Standard Methods for the Examination of Dairy Products, 1978, 14th Ed.,Washington D.C.
2.Atlas R. M., 2004, Handbook of Microbiological Media, 3rd Ed., CRC Press.
3.Diagnostic Procedures and Reagents, 1950, 3rd Edition, 13.
4.FDA Bacteriological Analytical Manual, 8th Ed., AOAC International, Gaithersburg, MD.
5.Huntoon F. M., 1918, J. Inf. Dis., 23:169.
6.Isenberg, H.D. Clinical Microbiology Procedures Handbook 2nd Edition.
7.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 8.Murray P. R., Baron J. H., Pfaller M. A., Jorgensen J. H. and Yolken R. H., (Eds.), 2003, Manual of Clinical Microbiology, 8th Ed., American Society for Microbiology, Washington, D.C. 9.Salfinger Y., and Tortorello M.L., 2015, Compendium of Methods for the Microbiological Examination of Foods,5th Ed., American Public Health Association, Washington, D.C.
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