SS Agar (Salmonella Shigella Agar) M108D

SS Agar (Salmonella Shigella Agar) is a differential selective media used for the isolation of Salmonella and some
Shigella species from pathological specimens, suspected foodstuffs etc.
Composition**
Ingredients Gms / Litre
Proteose peptone 5.000
Lactose 10.000
Bile salts mixture 8.500
Sodium citrate 8.500
Sodium thiosulphate 8.500
Ferric citrate 1.000
Brilliant green 0.00033
Neutral red 0.025
Agar 13.500
Final pH ( at 25°C) 7.0±0.2
**Formula adjusted, standardized to suit performance parameters

Directions
Suspend 60.00 grams in 1000 ml 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.
Proteose peptone, beef extract provide 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 centre 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 centres. Growth of Salmonella species is uninhibited and appears as colourless colonies with black
centres resulting from H 2S production. Shigella species also grow as colourless colonies which do not produce H2S.
It is recommended 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 (7).

Quality Control
Appearance
Light yellow to pink homogeneous free flowing powder
Gelling
Firm, comparable with 1.35% Agar gel

Please refer disclaimer Overleaf.

HiMedia Laboratories Technical Data

Colour and Clarity of prepared medium

Reddish orange coloured clear to slightly opalescent gel forms in Petri plates.
Reaction
Reaction of 6.0% w/v aqueous solution at 25°C. pH : 7.0±0.2
pH
6.80-7.20
Cultural Response
M108D: Cultural characteristics observed after an incubation at 35 - 37°C for 18 - 24 hours.
Organism Inoculum Growth Recovery Colour of
(CFU) colony
Escherichia coli ATCC 50-100 fair 20-30% pink with bile
25922 precipitate
Enterobacter aerogenes 50-100 fair 20-30% cream pink
ATCC 13048
Enterococcus faecalis ATCC 50-100 none-poor <=10% colourless
29212
Proteus mirabilis ATCC 50-100 fair-good 30-40% colourless,
25933 may have black
centre
Salmonella choleraesuis 50-100 good-luxuriant >=50% colourless with
ATCC 12011 black centre
Salmonella Typhi ATCC 50-100 good-luxuriant >=50% colourless with
6539 black centre
Salmonella Typhimurium 50-100 good-luxuriant >=50% colourless with
ATCC 14028 black centre
Salmonella Enteritidis ATCC 50-100 good-luxuriant >=50% colourless with
13076 black centre
Shigella flexneri ATCC 50-100 good 30-40% colourless
12022

Storage and Shelf Life

Store below 30°C in tightly closed container and the prepared medium at 2-8°C. Use before expiry date on the label.

Reference
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 Water
and 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 and
Wilkins, Baltimore.

Revision : 1 / 2011

Please refer disclaimer Overleaf.

HiMedia Laboratories Technical Data

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