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Technical Data
SS Agar (Salmonella Shigella Agar) M108
Intended Use:
Recommended for the isolation of Salmonella and some Shigella species from pathological specimens, suspected
foodstuffs etc.
Composition**
Ingredients g/L
Peptone 5.000
HM peptone B # 5.000
Lactose 10.000
Bile salts mixture 8.500
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
# - Equivalent to Beef extract
Directions
Suspend 63.02 grams in 1000 ml purified /distilled water. Boil with frequent agitation to dissolve the medium completely.
DO NOT AUTOCLAVE OR OVERHEAT. Overheating may destroy selectivity of the medium. Cool to about 50°C. Mix 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 (2,3,4,5) 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.
Peptone, HM peptone B provides nitrogen and carbon source, long chain amino acids, vitamins and essential growth
nutrients. Lactose is the fermentable carbohydrate. Brilliant green, bile salts and 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 Salmonella Shigella 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
Clinical: faeces, rectal swabs; Suspected food stuffs.
Specimen Collection and Handling
For clinical samples follow appropriate techniques for handling specimens as per established guidelines (8,9).
For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines (2,3,4,5).
After use, contaminated materials must be sterilized by autoclaving before discarding.

Warning and Precautions

In Vitro diagnostic use. For professional use only. 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 clinical specimens. Safety
guidelines may be referred in individual safety data sheets.

Please refer disclaimer Overleaf.

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HiMedia Laboratories Information For Use

Limitations
1.The medium is highly selective and may be toxic to certain Salmonella or Shigella species. Hence it is recommended
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).
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 coloured 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 Growth Recovery Colour of
(CFU) colony
# Klebsiella aerogenes 50-100 fair 20-30% cream pink
ATCC 13048 (00175*)
Escherichia coli fair 20-30% pink with bile
50-100
ATCC 25922 (00013*) precipitate

Salmonella Choleraesuis 50-100 good-luxuriant >=50% colourless with

ATCC 12011
black centre
Salmonella Typhi 50-100 good-luxuriant >=50% colourless with
ATCC 6539 black centre
Enterococcus faecalis 50-100 none-poor <=10% colourless
ATCC 29212 (00087*)
Proteus mirabilis 50-100 fair-good 30-40% colourless, may
ATCC 25933 have black centre
Shigella flexneri 50-100 good 40-50% colourless
ATCC 12022 (00126*)
Salmonella Typhimurium 50-100 good-luxuriant >=50% colourless with
ATCC 14028 (00031*) black centre
Salmonella Enteritidis 50-100 good-luxuriant >=50% colourless with
ATCC 13076 (00030*) 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 clinical sample
must be decontaminated and disposed of in accordance with current laboratory techniques (8,9).

Please refer disclaimer Overleaf.

HiMedia Laboratories Information For Use

Reference
1. Lennette and others (Eds.), 1985, Manual of Clinical Microbiology, 4th ed., ASM, Washington, D.C.
2. Lipps WC, Braun-Howland EB, Baxter TE,eds. Standard methods for the Examination of Water and Wastewater, 24th
ed. Washington DC:APHA Press; 2023.
3. Salfinger Y., and Tortorello M.L. Fifth (Ed.), 2015, Compendium of Methods for the Microbiological Examination of
Foods, 5th Ed., American Public Health Association, Washington, D.C.
4. Wehr H. M. and Frank J. H., 2004, Standard Methods for the Microbiological Examination of Dairy Products, 17th
Ed.,APHA Inc., 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-National Formulatory (USP-NF), 2022.
7. MacFaddin J., 1985, Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria, Vol. I, Williams
and Wilkins, 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
of Clinical Microbiology, 11th Edition. Vol. 1.

Revision : 05/2024

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Thane (W) -400604, MS, India 10°C

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Disclaimer :
User must ensure suitability of the product(s) in their application prior to use. Products conform solely to the information contained in this and
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of our knowledge true and accurate. HiMedia™ Laboratories Pvt Ltd reserves the right to make changes to specifications and information related
to the products at any time. Products are not intended for human or animal or therapeutic use but for laboratory,diagnostic, research or further
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