TRICHOPHYTON AGAR

Cat. no. C51 Trichophyton Agar #1, 20x125mm Tube, 10.5ml Slant 20 tubes/box
Cat. no. C52 Trichophyton Agar #2, 20x125mm Tube, 10.5ml Slant 20 tubes/box
Cat. no. C53 Trichophyton Agar #3, 20x125mm Tube, 10.5ml Slant 20 tubes/box
Cat. no. C54 Trichophyton Agar #4, 20x125mm Tube, 10.5ml Slant 20 tubes/box
Cat. no. C55 Trichophyton Agar #5, 20x125mm Tube, 10.5ml Slant 20 tubes/box
Cat. no. C56 Trichophyton Agar #6, 20x125mm Tube, 10.5ml Slant 20 tubes/box
Cat. no. C57 Trichophyton Agar #7, 20x125mm Tube, 10.5ml Slant 20 tubes/box

INTENDED USE

Hardy Diagnostics Trichophyton Media numbers 1-7 are used as an aid in the identification of various Trichophyton species.

SUMMARY

Some species of Trichophyton resemble one another closely and are difficult to differentiate on the basis of colony formation or microscopic morphology. In 1957, Georg and Camp formulated the Trichophyton Agars based on the nutritional requirements of the various Trichophyton species. These nutritional tests are used to aid in the identification of Trichophyton species.

FORMULA

Ingredients per liter of deionized water:*

Trichophyton Agar #1:
Casein Base Medium:
Dextrose 40.0gm
Casamino Acids 2.5gm
Monopotassium Phosphate 1.8gm
Magnesium Sulfate 0.1gm
Agar 15.0gm

Trichophyton Agar #2:
Casein Base Medium:
Dextrose 40.0gm
Casamino Acids 2.5gm
Monopotassium Phosphate 1.8gm
Magnesium Sulfate 0.1gm
Inositol 50.0mg
Agar 15.0gm

Trichophyton Agar #3:
Casein Base Medium:
Dextrose 40.0gm
Casamino Acids 2.5gm
Monopotassium Phosphate 1.8gm
Magnesium Sulfate 0.1gm
Inositol 0.50mg
Thiamine Hydrochloride, USP 200.0mcg
Agar 15.0gm

Trichophyton Agar #4:
Casein Base Medium:
Dextrose 40.0gm
Casamino Acids 2.5gm
Monopotassium Phosphate 1.8gm
Magnesium Sulfate 0.1gm
Thiamine Hydrochloride, USP 200.0mcg
Agar 15.0gm

Trichophyton Agar #5:
Casein Base Medium:
Dextrose 40.0gm
Casamino Acids 2.5gm
Monopotassium Phosphate 1.8gm
Magnesium Sulfate 0.1gm
Nicotinic Acid 2.0mg
Agar 15.0gm

Trichophyton Agar #6:
Ammonium Nitrate Agar Base:
Dextrose 40.0gm
Monopotassium Phosphate 1.8gm
Ammonium Nitrate 1.5gm
Magnesium Sulfate 0.1gm
Agar 15.0gm

Trichophyton Agar #7:
Ammonium Nitrate Agar Base:
Dextrose 40.0gm
Monopotassium Phosphate 1.8gm
Ammonium Nitrate 1.5gm
Magnesium Sulfate 0.1gm
Agar 15.0gm
L-Histidine HCl 0.03g/L

Final pH 6.8 +/- 0.2 at 25ºC.

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

STORAGE AND SHELF LIFE

Upon receipt store at 2-8ºC. away from direct light. Media should not be used if there are any signs of contamination, deterioration (shrinking, cracking, or discoloration), or if the expiration date has passed. Product is light and temperature sensitive; protect from light, excessive heat, moisture, and freezing.

PRECAUTIONS

PROCEDURE

Specimen Collection: Not applicable since these media are not primary isolation. These media are used in characterizing pure cultures. Isolated organisms, established isolation techniques, and tests for purity are necessary before inoculating these media. Direct inoculation of specimens will produce erroneous results. Information on specimen collection may be found in standard reference texts.

Method of Use: Prior to inoculation, the medium should be brought to room temperature. Place a small amount (about 1mm) of the test fungus on the surface of the test media. Be careful not to transfer agar from the media on which the test isolate is growing. Place a small amount of the test fungus on Sabouraud Dextrose Agar slant. Incubate the inoculated tubes for two weeks at room temperature (15-30ºC). If results are inconclusive at that time, reincubate the cultures for two more weeks or until the Sabouraud control tube shows good growth (up to 6 weeks). The tube in the test showing maximum growth is recorded as 4+ and other tubes are read in comparison. Examine each tube grossly and quantitate by comparison of the tubes. Growth on Trichophyton Agar # 2, 3, and 4 is compared to medium 1 to determine whether the isolate requires inositol, thiamine, or both. Trichophyton Agar 5 has added nicotinic acid and is compared to medium 1. Medium 7 has added histidine and is compared to medium 6.

4+ = maximum growth for the series of tests in comparison with colony growth of the other tests.
1+, 2+, 3+ = Trichophyton Agar read by comparison with the 4+ growth.
+/- = trace growth: results read by comparison to the 4+ growth.
0 = no growth.

INTERPRETATION OF RESULTS

Quantitate and compare growth with control tube. Consult appropriate references for growth patterns.

Typical Cultural Response of Certain Dermatophyte Species on Trichophyton Agars 1-7:
Species 1 2 3 4 5 6 7
Trichophyton verrucosum , 84% 0 +/- to 1+ 4+ 0





Trichophyton verrucosum , 16% 0 0 4+ 4+





Trichophyton schoenleinii 4+ 4+ 4+ 4+





Trichophyton concentricum , 50% 4+ 4+ 4+ 4+





Trichophyton concentricum , 50% 2+ 2+ 4+ 4+





Trichophyton violaceum +/- ND ND 4+





Trichophyton tonsurans +/- - to 1+ 3+ 4+





Trichophyton rubrum 4+ 4+ 4+ 4+





Trichophyton mentagrophytes 4+



4+





Trichophyton equinum 0





4+



Trichophyton megninii









0 4+
Microsporum gallinae









4+ 4+

LIMITATIONS

Pure cultures of mold must be grown on non-vitamin enriched media such as Sabouraud Dextrose Agar or Mycobiotic Agar prior to inoculation.

Cultures contaminated with bacteria must be repeatedly grown on a medium containing antimicrobials such as Mycobiotic Agar or Brain Heart Infusion (BHI) Agar with Cycloheximide and Chloramphenicol. Many bacteria synthesize vitamins which may invalidate the test.

When inoculating the Trichophyton Agars, it is important not to transfer growth substances from primary cultures to the tube media. The inoculum should be small.

Some Trichophyton spp. may require incubation at 35 +/- 2ºC. Consult listed references. (4)

MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as loops, other culture media, swabs, applicator sticks, 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
Trichophyton Agar #1-4
Trichophyton mentagrophytes
ATCC ® 9533
G 7 days 15-30°C Aerobic Growth; 3 - 4+
Trichophyton equinum
ATCC ® 22443
G 7 days 15-30°C Aerobic Inhibited; 0 - +/-
Trichophyton Agar #5
Trichophyton mentagrophytes
ATCC ® 9533
G 7 days 15-30°C Aerobic Growth; 3 - 4+
Trichophyton tonsurans
ATCC ® 28942
G 7 days 15-30°C Aerobic Inhibited; 0 - +/-
Trichophyton Agar #6
Trichophyton mentagrophytes
ATCC ® 9533
G 7-14 days 15-30°C Aerobic Growth; 4+
Trichophyton megninii
ATCC ® 12106
G
7-14 days 15-30°C Aerobic Inhibited; 0 - +/-
Trichophyton Agar #7
Trichophyton mentagrophytes
ATCC ® 9533
G 7-14 days 15-30°C Aerobic Growth; 2 - 4+
Trichophyton megninii
ATCC ® 12106
G 7-14 days 15-30°C Aerobic Growth; 2 - 4+

User Quality Control

PHYSICAL APPEARANCE

Trichophyton Agar products should appear light amber in color.

T. mentagrophytes growing on Trichophyton Agar

Trichophyton mentagrophytes (ATCC ® 9533) growing on Trichophyton Agar #1 (Cat. no. C51). Incubated aerobically for 7 days at 30ºC.

T. mentagrophytes growing on Trichophyton Agar

Trichophyton equinum (ATCC ® 22443) growth inhibited on Trichophyton Agar #1 (Cat. no. C51). Incubated aerobically for 7 days at 30ºC.

REFERENCES

1. Campbell, M.C. and J.L. Stewart. 1980. The Medical Mycology Handbook, John Wiley & Sons, New York, NY.

2. Georg, L.K. and L.B. Camp. 1957. J. Bacteriol.; 74:113.

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

4. Haley, L.D. and C.S. Calloway. 1984. Laboratory Methods in Medical Mycology, 4th ed. U.S. Department of Health and Welfare, Manual of Clinical Mycology, 5th ed. ASM, Washington, D.C.

5. Difco Manual, 10th ed. 1984. Difco Laboratories, Detroit, Michigan.

6. Hugh, R., and Leifson, J. 1957. J. Bacteriol.; 74:113.


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041816gr