SabHI AGAR

Cat. no. W75 SabHI Agar, 15x100mm Plate, 26ml 10 plates/bag
Cat. no. X75 SabHI Agar, 50ml HardyFlask™, 12ml 20 flasks/box
Cat. no. X73 SabHI Agar with Blood, Chloramphenicol
and Cycloheximide, 50ml HardyFlask™, 12ml
20 flasks/box

INTENDED USE

Hardy Diagnostics SabHI Agar is recommended for use in the cultivation of fungi. SabHI Agar with Blood, Chloramphenicol, and Cycloheximide is recommended for the selective isolation of pathogenic fungi.

SUMMARY

Sabouraud designed Sabouraud Dextrose Agar for the cultivation of dermatophytes.(9)  It is a general purpose medium that is used in qualitative procedures for the cultivation of dermatophytes and other pathogenic and non-pathogenic fungi from clinical and non-clinical specimens. Brain Heart Infusion Agar is used for the primary isolation and cultivation of fungi from clinical specimens.(2)  Gorman, in 1967, combined the two mediums to produce SabHI Agar. The combined formulation yields greater recovery of pathogenic fungi than either medium individually.(8)

The incorporation of chloramphenicol and cycloheximide provides selectivity to the medium. Chloramphenicol inhibits a range of gram-positive and gram-negative bacteria; cycloheximide inhibits saprophytic molds but may also inhibit growth of some significant pathogens (e.g., Cryptococcus neoformans, some Candida species, some Aspergillus spp. and mucormycetes (formally zygomycetes)). Blood provides essential growth factors for the more fastidious fungal organisms. Gorman showed that the addition of blood increased the recovery of H. capsulatum.(8) Blood also aids in the conversion of H. capsulatum and B. dermatitidis to the yeast phase.

FORMULA

Ingredients per liter of deionized water:*

Dextrose 21.0gm
Brain Heart Infusion 10.0gm
Meat Peptone 7.25gm
Sodium Chloride 2.5gm
Casein Peptone 2.25gm
Disodium Phosphate 1.25gm
Agar 15.0gm

Additionally, SabHI Agar with Blood, Chloramphenicol and Cycloheximide contains:

Cycloheximide 500.0mg
Chloramphenicol 50.0mg
Sheep Blood 50.0ml

Final pH 7.0 +/- 0.3 at 25ºC.

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

STORAGE AND SHELF LIFE

Storage: Upon receipt store Cat. nos. W75 and X73 at 2-8ºC away from direct light. Store Cat. no. X75 at 2-30ºC away from direct light. Media should not be used if there are any signs of deterioration (shrinking, cracking, or discoloration), hemolysis, contamination, or if the expiration date has passed.

PRECAUTIONS

PROCEDURE

Specimen Collection: Infectious material should be submitted directly to the laboratory without delay and protected from excessive heat and cold. If there is to be a delay in processing, the specimen should be inoculated onto an appropriate transport media and refrigerated until inoculation. Consult listed references for information regarding the processing and inoculation of specimens.(2-6)

Method of Use: Inoculate a non-selective and selective medium in parallel to ensure recovery of pathogenic fungi from potentially contaminated specimens. Incubate aerobically at 25-30ºC with increased humidity for 30 days or longer. Inoculate two sets of media for isolation of fungi that cause systemic mycoses; incubate one set at 25-30ºC and the other set at 35 +/- 2ºC . Examine cultures weekly for a period of four weeks before being reported negative.

INTERPRETATION OF RESULTS

Media should be examined for characteristic colonial growth and morphology. Consult listed references for the interpretation of fungal growth on this medium.(2-6)

LIMITATIONS

For proper identification of fungi, microscopic examination and evaluation of morphological structures is required.

For selective media, specific strains of fungi for which the medium is designed to isolate often may be inhibited. Fungi for which the medium is designed to inhibit may grow.

A non-selective and selective medium should be inoculated for isolation of fungi from potentially contaminated specimens.

MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as loops, swabs, applicator sticks, other culture media, incinerators, incubators, etc., as well as serological and biochemical reagents, are not provided.

QUALITY CONTROL

Test Organisms Inoculation Method* Incubation Results
Time Temperature Atmosphere
SabHI Agar:
Trichophyton mentagrophytes
ATCC ® 9533**
G 7 days 15-30°C Aerobic Growth
Candida albicans
ATCC ® 60193**
A 24-48hr 35°C Aerobic Growth
SabHI Agar with Blood, Chloramphenicol and Cycloheximide:
Candida albicans
ATCC ® 10231**
A 48hr 35°C Aerobic Growth
Trichophyton mentagrophytes
ATCC ® 9533**
G 7 days 15-30°C Aerobic Growth
Aspergillus brasiliensis
ATCC ® 16404
G 7 days 15-30°C Aerobic Partial to complete inhibition
Escherichia coli
ATCC ® 25922**
B 24hr 35°C Aerobic Partial to complete inhibition

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

User Quality Control

PHYSICAL APPEARANCE

SabHI Agar with and without antimicrobics, should appear clear, and amber in color.

SabHI Agar with Blood, Chloramphenicol and Cycloheximide should appear opaque, and cherry red in color.

T. mentagrophytes growing on SabHI Agar

Trichophyton mentagrophytes (ATCC ® 9533) growing on SabHI Agar (Cat. no. W75). Incubated aerobically for 72 hours at 30ºC .

C. albicans growing on SabHI Agar

Candida albicans (ATCC ® 60193) colonies growing on SabHI Agar (Cat. no. W75). Incubated aerobically for 24 hours at 35ºC .



SabHI Agar

Uninoculated plate of SabHI Agar (Cat. no. W75).



REFERENCES

1. Anderson, N.L., et al. Cumitech 3B; Quality Systems in the Clinical Microbiology Laboratory, Coordinating ed., A.S. Weissfeld. American Society for Microbiology, Washington, D.C.

2. Jorgensen., et al. Manual of Clinical Microbiology, American Society for Microbiology, Washington, D.C.

3. Tille, P., et al. Bailey and Scott's Diagnostic Microbiology, C.V. Mosby Company, St. Louis, MO.

4. Cumitech 11; Practical Methods for Culture and Identification of Fungi in the Clinical Microbiology Laboratory , American Society for Microbiology, Washington, D.C., 1980.

5. Isenberg, H.D. Clinical Microbiology Procedures Handbook, Vol. I, II & III. American Society for Microbiology, Washington, D.C.

6. Koneman, E.W., et al. Color Atlas and Textbook of Diagnostic Microbiology, 6th ed. J.B. Lippincott Company, Philadelphia, PA.

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

8. Gorman, J.W. 1967. Am. J. Med. Technol.; 33:151.

9. Sabouraud. 1892. Ann. Dermatol. Syphil.; 3:1061.


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

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