SIM HiCynth™ Medium

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MCD181
Recommended for determination of hydrogen sulphide production, indole formation and motility of enteric bacilli from clinical and non-clinical samples.


SIM HiCynth™ Medium is recommended for determination of hydrogen sulphide production, indole formation and motility of enteric bacilli.

Composition**

Ingredients Gms / Litre
HCynth™ Peptone No.1* 33.000
Iron Substrate 0.230
Sodium thiosulphate 0.025
Agar 3.000
Final pH (at 25°C) 7.3±0.2

**Formula adjusted, standardized to suit performance parameters
*Chemically defined peptone

Directions

Suspend 36.23 grams in 1000 ml distilled water. Heat to boiling to dissolve the medium completely. Dispense in tubes. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Allow the tubes to cool in an upright position.

Principle And Interpretation

SIM HiCynth™ Medium is used to differentiate enteric bacilli particularly Salmonella and Shigella on the basis of sulphide production, indole formation and motility (1,2). Jordan and Victorson (3) reported that Salmonella Paratyphi A and Paratyphi B can be distinguished on the basis of H2S production using lead acetate. Sulkin and Willett (4) used Triple Sugar Iron Agar with 1% agar for motility along with H2S production and carbohydrate fermentation. Sosa (5) described a medium with low agar for motility and indole determination.

SIM HiCynth™ Medium enables determination of three characteristics by which enteric bacteria can be differentiated. H2S production is indicated by black precipitate of ferrous sulphide. Motile organisms intensify the H2S reaction. Motile organisms grow away from line of inoculation showing diffused growth while non-motile organisms grow along the stabline. Motility detection is possible due to the semisolid nature of the medium. Growth radiating out from the central stabline indicates that the test organism is motile.

HiCynth™ Peptone No. 1 provides nitrogeneous and carbonaceous compounds, long chain amino acids, vitamins and other essential growth nutrients. Tryptophan, from the medium, is degraded by specific bacteria to produce indole (2). The indole is detected by the addition of Kovacs reagents following the incubation period.

Inoculate fresh culture with a single stab using straight needle through the center of the medium. Following incubation, observe for motility (diffuse growth outward from the stabline or turbidity throughout the medium) and for H2S production (blackening of the medium). To detect indole production, add three or four drops of Kovacs reagent (2) and observe for development of red color (positive reaction). Determine motility and H2S production prior to determination of indole production.

Quality Control

Appearance
Cream to beige homogeneous free flowing powder

Gelling
Semisolid, comparable with 0.3% Agar gel.

Colour and Clarity of prepared medium
Medium amber coloured slightly opalescent gel forms in tubes as butts

Reaction
Reaction of 3.6% w/v aqueous solution at 25°C. pH: 7.3±0.2

Cultural Response

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

Cultural Response

Organism Inoculum (CFU) Growth Motility Indole H2S production(on addition of Kovac's Reagent)
Escherichia coli ATCC 25922 50-100 luxuriant positive, growth away from stabline causing turbidity positive reaction, red ring at the interface of the medium negative reaction
Salmonella Typhimurium ATCC 14028 50-100 luxuriant positive, growth away from stabline causing turbidity negative reaction positive reaction, blackening of medium
Shigella flexneri ATCC 12022 50-100 luxuriant negative, growth along the stabline, surrounding medium remains clear negative reaction negative reaction
Salmonella Paratyphi A ATCC 9150 50-100 luxuriant positive, growth away from stabline causing turbidity negative reaction Negative reaction
Salmonella Paratyphi B ATCC 8739 50-100 luxuriant positive, growth away from stabline causing turbidity Negative reaction Positive reaction, blackening of medium
Klebsiella pneumoniae ATCC 13883 50-100 luxuriant negative, growth along the stabline, surrounding medium remains clear Negative reaction Negative reaction

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.

More Information
Product Name SIM HiCynth™ Medium
SKU MCD181
Product Type HiCynth™
Physical Form Powder
Origin Chemically defined (HiCynth™)
Packaging type HDPE
References 1. MacFaddin J. F., 1985, Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria, Vol. 1, Williamsand Wilkins, Baltimore.
2.Ewing W. H., 1986, Edwards and Ewings Identification of Enterobacteriaceae, 4th Ed., Elsevier Science Publishing Co.,Inc. New York.
3.Jordan E. O. and Victorson R., 1917, J. Inf. Dis., 21:554.
4.Sulkin S. E. and Willett J. C., 1940, J. Lab. Clin. Med., 25:649.
5.Sosa L., 1943, Rev. Inst. Bacteriol., 11:286.
6.Isenberg, H.D. Clinical Microbiology Procedures Handb0ook. 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.
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