Mannitol Salt HiVeg™ Agar Base

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MV118
For selective isolation of pathogenic Staphylococci.


Intended Use

Recommended for selective isolation of pathogenic Staphylococci from various samples.

Composition**

Ingredients g/L
HiVeg® peptone No. 3 10.000
HiVeg® extract 1.000
Sodium chloride 75.000
D-Mannitol 10.000
Phenol red 0.025
Agar 15.000
Final pH (at 25°C) 7.4±0.2

**Formula adjusted, standardized to suit performance parameters

Directions

Suspend 111.02 grams in 1000 ml 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, add PRTC Selective Supplement (FD045). Mix well and pour into sterile Petri plates.

Note : This product contains 7.5% Sodium chloride as one of its ingredients. On repeated exposure to air and absorption moisture, sodium chloride has tendency to form lumps, therefore we strongly recommend storage in tightly closed containers in dry place away from bright light.

Principle And Interpretation

Staphylococci are widespread in nature, although they are mainly found on the skin, skin glands and mucous membranes of mammals and birds. The coagulase-positive species i.e Staphylococcus aureus is well documented as a human opportunistic pathogen. The ability to clot plasma continues to be the most widely used and accepted criterion for the identification of pathogenic staphylococci associated with acute infections (1). Staphylococci have the unique ability of growing on a high salt containing media (2). Isolation of coagulase-positive Staphylococci on Phenol Red Mannitol Agar supplemented with 7.5%NaCl was studied by Chapman (3). The resulting Mannitol Salt Agar Base is recommended for the isolation of coagulase-positive Staphylococci from cosmetics, milk, food and other specimens (1,4,5,6,7). The additional property of lipase activity of Staphylococcus aureus can be detected by the addition of the Egg Yolk Emulsion (FD045). The lipase activity can be visualized as yellow opaque zones around the colonies (2).

Mannitol Salt HiVeg® Agar Base is same as Mannitol Salt Agar Base except that the animal based peptones are completely replaced with vegetable peptones to avoid the BSE/TSE risks associated with animal peptones.

HiVeg® peptone No. 3 and HiVeg® extract supply essential growth factors and trace nutrients to the growing bacteria. Sodium chloride serves as an inhibitory agent against bacteria other than Staphylococci. Mannitol is the fermentable carbohydrate, fermentation of which leads to acid production, detected by phenol red indicator. S.aureus ferments mannitol and produce yellow coloured colonies surrounded by yellow zones. Coagulase-negative strains of S.aureus are usually mannitol non-fermenters and therefore produce pink to red colonies surrounded by red-purple zones. Presumptive coagulase-positive yellow colonies of S. aureus should be confirmed by performing the coagulase test [tube or slide] (1). Lipase activity of S.aureus can be detected by supplementing the medium with egg yolk emulsion.

A possible S.aureus must be confirmed by the coagulase test. Also the organism should be subcultured to a less inhibitory medium not containing excess salt to avoid the possible interference of salt with coagulase testing or other diagnostic tests (e.g. Nutrient HiVeg® Broth) (MV002) (8). Few strains of S.aureus may exhibit delayed mannitol fermentation. Negative results should therefore be re-incubated for an additional 24 hours before being discarded (8).

Type of specimen

Food and dairy samples, Water samples.

Specimen Collection and Handling

For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines (6,9). 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. A possible S.aureus must be confirmed by the coagulase test.
  2. The organism should be subcultured to a less inhibitory medium not containing excess salt to avoid the possible interference of salt with coagulase testing or other diagnostic tests (e.g. Nutrient HiVeg™ Broth) (MV002).
  3. Few strains of S.aureus may exhibit delayed mannitol fermentation. Negative results should therefore be re-incubated for an additional 24 hours before being discarded.

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
Light yellow to pink homogeneous free flowing powder

Gelling
Firm,comparable with 1.5% Agar gel

Colour and Clarity of prepared medium
Red coloured clear to slightly opalescent gel forms in Petri plates

Reaction
Reaction of 11.1% w/v aqueous solution at 25°C. pH: 7.4±0.2

pH
7.20-7.60

Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 18-72 hours. Recovery rate is considered as 100% for bacteria growth on Soybean Casein Digest Agar.

Organism Inoculum (CFU) Growth Recovery Colour of colony
Staphylococcus aureus subsp. aureus ATCC 6538 (00032*) 50-100 luxuriant >=50% yellow/white colonies surrounded by yellow zone
Escherichia coli ATCC 8739 (00012*) >=104 inhibited 0%
Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) 50-100 luxuriant >=50% yellow/white colonies surrounded by yellow zone
Staphylococcus epidermidis ATCC 14990 (00132*) 50-100 fair-good 30-40% red
Proteus mirabilis ATCC 12453 50-100 none-poor 0-10% yellow
Escherichia coli ATCC 25922 (00013*) >=104 inhibited 0%
# Klebsiella aerogenes ATCC 13048 (00175*) >=104 inhibited 0%

Key

Key: (*) Corresponding WDCM numbers. # - Formerly known as Enterobacter aerogenes

Storage and Shelf Life

Store between 10-30°C in a tightly closed container and the prepared medium at 20-30°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 (10,11).

More Information
Product Name Mannitol Salt HiVeg™ Agar Base
SKU MV118
Product Type HiVeg™
Physical Form Powder
Origin Animal Free (Veg)
Packaging type HDPE
References 1.Murray P. R., Baron J. H., Pfaller M. A., Jorgensen J. H. and Yolken R. H., (Ed.), 2003, Manual of Clinical Microbiology,8th Ed., American Society for Microbiology, Washington, D.C. ,,2.Koch P. K., 1942, Zentralbl. Bakteriol. Parasitenkd. Abt. I Orig.149:122.
3.Chapman G. H., 1945, J. Bacteriol., 50:201.
4.Hitchins A. D., Tran T. and McCarron J. E., 1995, FDA Bacteriological Analytical Manual, 8th Ed., AOAC International,Gaithersburg, Md.
5.Davis J. G., 1959, Milk testing, 2nd Ed., Dairy Industries Ltd, London.
6.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.
7.Silverton R. E. and Anderson M. J., 1961, Handbook of Medical Laboratory Formulae, Butterworths, London.
8.Gunn B. A., Dunkelberg W. E. and Creitz J. R., 1972, Am. J. Clin. Pathol., 57:236.9.MacFaddin J. F., 1985, Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria, Vol. 1, Williamsand Wilkins, Baltimore10.Isenberg, H.D. Clinical Microbiology Procedures Handbook. 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. 1.1
2.American Public Health Association, Standard Methods for the Examination of Dairy Products, 1978, 14th Ed., WashingtonD.C.
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