SS HiVeg™ Agar (Salmonella Shigella HiVeg™ Agar)

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SKU:
MV108
For differential and selective isolation of Salmonella and Shigella species from pathological specimens, suspected foodtuffs etc.


Intended Use

Recommended as a differential selective media for isolation of Salmonella and some Shigella species from various samples.

Composition**

Ingredients g/L
HiVeg™ peptone 11.500
HiVeg™ extract 5.000
Lactose 10.000
Synthetic detergent No. I 2.000
Sodium citrate 10.000
Sodium thiosulphate 8.500
Ferric citrate 1.000
Brilliant green 0.00033
Neutral red 0.025
Agar 15.000
Final pH (at 25°C) 7.0±0.2

**Formula adjusted, standardized to suit performance parameters

Directions

Suspend 63.02 grams in 1000 ml purified / distilled water. Heat to boiling with frequent agitation to dissolve the medium completely. DO NOT AUTOCLAVE OR OVERHEAT. Overheating may destroy selectivity of the medium. Cool to 45-50°C. Mix well and pour into sterile Petri plates.

Principle And Interpretation

SS Agar medium is recommended as differential and selective medium for the isolation of Salmonella and Shigella species from pathological specimens (1) and suspected foodstuffs (2-5) and for microbial limit test (6). SS Agar is a moderately selective medium in which gram-positive bacteria are inhibited by bile salts, brilliant green and sodium citrate. SS HiVeg™ Agar is same as SS Agar except that the animal based peptones are completely replaced with vegetable peptones to avoid the BSE/TSE risks associated with animal peptones.

HiVeg™ peptone, HiVeg™ extract provides nitrogen and carbon source, long chain amino acids, vitamins and essential growth nutrients. Lactose is the fermentable carbohydrate. Brilliant green, synthetic detergent No. I and sodium thiosulphate selectively inhibit gram-positive and coliform organisms. Sodium thiosulphate is reduced by certain species of enteric organisms to sulphite and H2S gas and this reductive enzyme process is attributed by thiosulphate reductase. Production of H2S gas is detected as an insoluble black precipitate of ferrous sulphide, formed upon reaction of H2S with ferric ions or ferric citrate, indicated in the center of the colonies.

The high selectivity of SS HiVeg™ Agar allows the use of large inocula directly from faeces, rectal swabs or other materials suspected of containing pathogenic enteric bacilli. On fermentation of lactose by few lactose-fermenting normal intestinal flora, acid is produced which is indicated by change of colour from yellow to red by the pH indicator- neutral red. Thus these organisms grow as red pigmented colonies. Lactose non-fermenting organisms grow as translucent colourless colonies with or without black centers. Growth of Salmonella species appears as colourless colonies with black centers resulting from H2S production. Shigella species also grow as colourless colonies which do not produce H2S.

Type of specimen

Suspected food stuffs.

Specimen Collection and Handling

For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines (2-5).

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. The medium is highly selective and may be toxic to certain Salmonella or Shigella species. Hence it isrecommended to use to inoculate plates of less inhibitory media parallel to SS Agar, such as Hektoen Enteric Agar (M467) or Deoxycholate Citrate Agar (M065) for easier isolation of Shigella species (9).
  2. Further biochemical and serological tests need to be carried out for confirmation.

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
Reddish orange coloured clear to slightly opalescent gel forms in Petri plates

Reaction
Reaction of 6.3% w/v aqueous solution at 25°C. pH: 7.0±0.2

pH
6.80-7.20

Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 18-24 hours.

Organism Inoculum
(CFU)
Growth Recovery Colour of
colony
# Klebsiella aerogenes
ATCC 13048 (00175*)
50-100 fair 20-30% cream pink
Escherichia coli ATCC
25922 (00013*)
50-100 fair 20-30% pink with bile
precipitate
Salmonella Choleraesuis
ATCC 12011
50-100 good-luxuriant >=50% colourless with
black centre
Salmonella Typhi
ATCC 6539
50-100 good-luxuriant >=50% colourless with
black centre
Enterococcus faecalis
ATCC 29212 (00087*)
50-100 none-poor <=10% colourless
Proteus mirabilis
ATCC 25933
50-100 fair-good 30-40% colourless, may
have black
centre
Shigella flexneri
ATCC 12022 (00126*)
50-100 good 40-50% colourless
Salmonella Typhimurium
ATCC 14028 (00031*)
50-100 good-luxuriant >=50% colourless with
black centre
Salmonella Enteritidis
ATCC 13076 (00030*)
50-100 good-luxuriant >=50% colourless with
black centre

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 (8,9).

More Information
Product Name SS HiVeg™ Agar (Salmonella Shigella HiVeg™ Agar)
SKU MV108
Product Type HiVeg™
Physical Form Powder
Origin Animal Free (Veg), Lactose
Packaging type HDPE
References 1.Lennette and others (Eds.), 1985, Manual of Clinical Microbiology, 4th ed., ASM, Washington, D.C.
2.Downes F. P. and Ito K., (Eds.), 2001, Compendium of Methods for the Microbiological Examination of Foods, 4th Ed.,APHA, Washington, D.C.
3.Wehr H. M. and Frank J. H., 2004, Standard Methods for the Microbiological Examination of Dairy Products, 17th Ed.,APHA Inc., Washington, D.C.
4.Eaton A. D., Clesceri L. S., Rice E. W., and Greenberg A. W., (Eds.), 2005, Standard Methods for the Examination of Waterand Wastewater, 21st Ed., APHA, Washington, D.C.
5.Williams S., (Ed.), 2005, Official Methods of Analysis of the Association of Official Analytical Chemists, 19th Ed., AOAC,Washington, D.C.
6.The United States Pharmacopoeia, 2006, USP29/NF24, The United States Pharmacopoeial Convention. Rockville, MD.
7.MacFaddin J., 1985, Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria, Vol. I, Williams andWilkins, Baltimore. 8.Isenberg, H.D. Clinical Microbiology Procedures Handbook. 2nd Edition.
9.Jorgensen,J.H., Pfaller , M.A., Carroll, K.C., Funke, G., Landry, M.L., Richter, S.S and Warnock., D.W. (2015) Manual ofClinical Microbiology, 11th Edition. Vol. 1.
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