m staphylococcus Broth

Cat. no. K213 m Staphylococcus Broth, 16x125mm Tube, 10ml 20 tubes/box

INTENDED USE

Hardy Diagnostics m Staphylococcus Broth is recommended for the isolation and differentiation of Staphylococcus spp. by membrane filtration.

This product is not intended to be used for the diagnosis of human disease.

SUMMARY

The presence of staphylococci, along with other organisms, are indicators of recreational water quality. (1) Pseudomonas , Streptococcus , and Staphylococcus are normal skin flora that are likely to be shed and are indicators of health risk. (2) These organisms account for a large percentage of swimming pool-associated illness. (1) Staphylococci are widespread in nature, though they are mainly found living on the skin, skin glands, and mucous membranes of mammals and birds. (3) m Staphylococcus Broth is patterned after the formula of Staphylococcus Medium 110 that Chapman created while developing a selective medium for staphylococci. (4) For the multiple-tube procedure to monitor swimming pool water for S. aureus , sodium azide is added to m Staphylococcus Broth. (1)

m Staphylococcus Broth contains peptone to provide the nitrogen, amino acids, and minerals. Yeast extract provides the vitamin source. Lactose and mannitol are carbohydrates for bacterial growth. Dipotassium phosphate is the buffering agent and the high concentration of sodium chloride causes the medium to be selective for staphylococci.

FORMULA

Ingredients per liter of deionized water:*

Sodium Chloride 75.0gm
Pancreatic Digest of Casein 10.0gm
Mannitol 10.0gm
Dipotassium Phosphate 5.0gm
Yeast Extract 2.5gm
Lactose 2.0gm

Final pH 7.0 +/- 0.2 at 25ºC.

* Adjusted and/or supplemented as required to meet performance criteria.

STORAGE AND SHELF LIFE

Storage: Upon receipt store at 2-30ºC. away from direct light. Media should not be used if there are any signs of deterioration, discoloration, contamination, or if the expiration date has passed. Product is light and temperature sensitive; protect from light, excessive heat, moisture, and freezing.

PRECAUTIONS

PROCEDURE

1. Refer to Standard Methods for the Examination of Water and Wastewater for information on water sample collection, processing, and the membrane filtration procedure. (1)

2. Use 2.0 to 2.5ml of m Staphylococcus Broth to saturate an absorbant pad (Cat. no. B200G047A).

3. After the sample has been filtered, remove the membrane from the filtering apparatus and place the membrane on the saturated absorbant pad that was prepared in Step 2.

4. Incubate at 35ºC. for 40-48 hours under aerobic conditions.

INTERPRETATION OF RESULTS

Following incubation, examine the membrane for growth and pigment production. Mannitol fermentation can be tested for by adding a drop of bromothymol blue to the site from which a colony is removed; a yellow color indicates mannitol fermentation.

LIMITATIONS

Follow recommended procedures for confirming growth on the membranes. (1,5,6)

MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as loops, swabs, filters, applicator sticks, absorbant pads (Cat. no. B200G047A), Lipovitellin Salt Mannitol Agar (Cat. no. G206), other culture media, incinerators, and incubators, etc., as well as serological and biochemical reagents, are not provided.

QUALITY CONTROL

Test Organisms Inoculation Method* Incubation Results
Time Temperature Atmosphere
Staphylococcus aureus
ATCC ® 25923**
A 24hr 35°C Aerobic Growth; yellow colonies when subcultured to Lipovitellin Salt Mannitol Agar (Cat. no. G206)
Staphylococcus epidermidis
ATCC ® 12228
A 24hr 35°C Aerobic Growth; pink colonies when subcultured to Lipovitellin Salt Mannitol Agar (Cat. no. G206)
Escherichia coli
ATCC ® 25922**
B 24hr 35°C Aerobic Inhibited; no growth when subcultured to Lipovitellin Salt Mannitol Agar (Cat. no. G206)

** Recommended QC strains for User Quality Control according to the CLSI document M22 when applicable.

USER QUALITY CONTROL

Physical Appearance

m Staphylococcus Broth should appear clear to slightly opalescent, and light amber in color. May have a slight precipitate but no debris.

REFERENCES

1. American Public Health Association. Standard Methods for the Examination of Water and Wastewater, APHA, Washington, D.C.

2. Seyfried, Tobin, Brown and Ness. 1985. Am. J. Public Health ; 75:1071.

3. Kloos and Bannerman. 1999. In Murray, Baron, Pfaller, Tenover and Yolken (ed.), Manual of Clinical Microbiology , 7th ed. American Society for Microbiology, Washington, D.C.

4. Chapman. 1946. J. Bacteriol. ; 51:409.

5. Quality Assurance for Commercially Prepared Microbiological Culture Media , M22. Clinical and Laboratory Standards Institute (CLSI - formerly NCCLS), Wayne, PA.

6. Association of Official Analytical Chemists. Official Methods of Analysis , AOAC, Washington, D.C.


ATCC is a registered trademark of the American Type Culture Collection.


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