TECHNICAL DATA SHEET #755 REV.

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27120 SW 95th Avenue Wilsonville, OR 97070 800.547.0659
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TRICHOMONAS MEDIA
PRODUCTS:
Tubed Media:
Trichomonas Media, Feinberg T6399
Trichomonas Media, Diamonds Modied T3670, T6650
Trichomonas Media, Diamonds Broth T6658
Trichomonas Media, Kupferberg T6652
PURPOSE:
Trichomonas media are recommended for the detection and cultivation of Trichomonas species.
PRINCIPLE:
The superiority of the culture procedure over the wet mount procedure for detecting the presence of trichomonads in clinical
specimens was demonstrated by Williams,
10
and Kean and Day.
5
Feinberg and Whittington,
2
after a series of investigations,
demonstrated the greater accuracy of the culture procedure for detecting trichomonas organisms in clinical material and
stressed that negative cultures are the best criteria for ascertaining the efcacy of therapy in these infections. Feingberg media
was subsequently developed to obtain good growth of both Trichomonas sp. and Candida sp. in mixed culture. The growth of
Candida sp. does not interfere with that of Trichomonas. The Feinberg formulation over the years has been slightly modied by
the incorporation of 0.1% w/v of agar which leads to reduced oxygen tension and consequently a much more prolic growth of
trichomonads.
Diamonds Broth was developed to study trichmonads in animals and humans. The addition of antimicrobials helps supress
microorganisms that would otherwise ourish in such rich media. In 1989 Rashad et al.
9
studied the recovery of Trichomonas
from vaginal secretions and found that culturing the organism in Diamonds Modied media led to an increase in recovery of 22%
over the wet-mount method and a 37% increase in recovery over culturing using Feinberg media.
The Kupferberg
6,7
formula contains chloramphenicol which inhibits a wide range of gram-negative and gram-positive bacteria.
The addition of methylene blue acts as an indictor to show oxygen diffusion in the media.
FORMULAS:
Approximate, per liter deionized ltered water.
(1) Trichomonas Media, Feinberg:
Liver Digest .....................................................25.0 g
Sodium Chloride ...................................................6.5
Dextrose ...............................................................5.0
Agar ......................................................................1.5
Horse Serum ......................................................80.0 ml
Penicillin .................................................1,000,000.0 units
Streptomycin .............................................500,000.0
Final pH 6.5 0.2 at 25C
(2) Trichomonas Media, Diamonds Broth:
Pancreatic Digest of Casein ................................20.0 g
Yeast Extract .......................................................10.0
Maltose ................................................................. 5.0
Dipotassium Phosphate ....................................... 0.8
Monopotassium Phosphate .................................. 0.8
Agar ...................................................................... 0.5
L-Cysteine HCl...................................................... 1.0
L-Ascorbic Acid ..................................................... 0.2
Gentamicin ..........................................................20.0
Vancomycin .........................................................20.0
Horse Serum .....................................................100.0 ml
Final pH 6.0 0.2 at 25C
TECHNICAL DATA SHEET #755 REV. 3
27120 SW 95th Avenue Wilsonville, OR 97070 800.547.0659
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(3) Trichomonas Media, Diamonds Modied:
Same as (2) with the addition of 10.0 mg of Amphotericin B
(4) Trichomonas Media, Kupferberg:
Pancreatic Digest of Casein ................................10.0 g
Peptic Digest of Animal tissue .............................10.0
Maltose ................................................................. 1.0
Agar ...................................................................... 1.0
L-Cysteine HCl...................................................... 1.5
Chloroamphenicol .................................................0.1
Methylene Blue .................................................0.003
Final pH 6.0 0.2 at 25C
PRECAUTIONS:*
For in vitro diagnostic use. Observe approved biohazard precautions.
Storage: Upon receipt store at 2-8C away from direct light. Media should not be used if there are signs of contamination,
deterioration (evaporation or discoloration), or if the expiration date has passed.
Limitations: Selective media often inhibit, to some extent, the specic strains of organisms that they are designed to isolate,
while other organisms resistant to the concentration of antimicrobics used in the media may grow.
Trichomonads are adversely affected by oxygen; avoid aeration of the media before and after inoculation and inoculate to the
bottom of the tube.
PROCEDURE:*
Specimen Collection: Information on specimen collection is found in standard reference material on the subject.
4
In general,
specimens should be protected from extreme heat and cold and delivered to the laboratory without delay.
Method of Use: Prior to inoculation the media should be brought up to 35-37C in an aerobic incubator for approximately
1-2 hours. If media is to be inoculated at the bedside, then the media may be warmed by holding the tube under the arm or in the
hands prior to inoculation. T. vaginalis does not survive for very long in clinical specimens (i.e. genital and urine) and should be
inoculated into culture media as soon as possible to maximize recovery of the organism. Direct wet mounts of freshly collected
specimens may be examined for T. vaginalis prior to inoculation into culture media. Urine specimens should be concentrated by
centrifugation (10 minutes at 250 x g) and the sediment inoculated into culture media and/or examined microscopically. Tubes
may be incubated aerobically in a slanted position (45 angle) at 35-37C.
7
Cultures may be examined by wet mount for motile
trophozoites after 24 or 48 hours, and centrifugation of the culture may be used to detect low-level growth. During incubation,
the initial culture tube may be subcultured by dispersing the inoculum (shaking or vortexing) and transferring 1-2 ml into a warm,
fresh tube. Negative cultures should be incubated for at least 96 hours and examined before reporting the culture as negative.
It is suggested to culture the patient 1 week after the completion of therapy. If this culture is negative, a second culture is taken
in 2 to 4 weeks, and if negative, the patient is considered cured.
Interpretation: After incubation, a drop of the media is placed on the slide and coverslipped. If the examination under low power
magnication results in observing motile trophozoites, the test is considered positive for Trichomonas.
Materials Required but Not Provided: Standard microbiological supplies and equipment are not provided.
QUALITY CONTROL:*
Microorganisms Used (ATCC #): Expected Results:
Feinberg Diamonds, Modied and Kupferberg
Trichomonas vaginalis (30001) Growth Growth
Candida albicans (10231) Growth Inhibition, partial to complete
Staphylococcus aureus (25923) Inhibition, partial to complete Inhibition, partial to complete

Diamonds Broth
Trichomonas vaginalis (30001) Growth
Escherichia coli (25922) Inhibition, partial to complete
Key: See Interpretation
TECHNICAL DATA SHEET #755 REV. 3
27120 SW 95th Avenue Wilsonville, OR 97070 800.547.0659
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User Quality Control: Check for signs of contamination and deterioration. Feinberg media should appear hazy and brown in
color. Diamonds Broth and Diamonds Modied media should appear hazy and light yellow in color. Kupferberg media should
appear hazy and light yellow in color with a blue-green ring at the top of the media.
BIBLIOGRAPHY:
1. Diamond, L.S. The establishment of various trichonads of animals and man in axenic cultures. J. Parasit. 43: 488-490,
1957.
2. Feinberg, J . G., and J . M. Whittington, J. Clin. Pathol., 10:327-329, 1957.
3. Fouts, A. C., and S. J . Kraus, J. Infect. Dis., 141:137-143, 1980.
4. Isenberg, H. D., Clinical Microbiology Procedures Handbook, Vol. 2, ASM, Washington, D. C., 1992.
5. Kean, B. H., and E. Day, Am. J. Obst. and Gyn., 68:1510-1518, 1954.
6. Kupferberg, A.B., et al., Nutritional requirements for Trichomonas vaginalis. Proc. Soc. Exp. Bio. Med. 67:304-308, 1948.
7. Kupferberg, A.B., Trichomonas vaginalis: Nutritional requirements and Diagnostic Procedures. Int'l. Rec. Med. and Gen.
Pract. Clin. 168: 709-717, 1955.
8. Lennette, E. H., et al., Manual of Clinical Microbiology, 4th ed., American Society for Microbiology, Washington, D. C.,
1985.
9. Rashad, A. L., Interscience Conference on Antimicrobial Agents and Chemotherapy, Abstracts of the Annual Meeting, No.
517, Houston, 1989, p. 187.
10. Williams, M. H., Am. J. Obst. and Gyn., 60:224-225, 1950.
*For more detailed information, consult appropriate references.
bioMrieux, Inc.
Data #755
Revision Date: April 2009