trichomonas media

TECHNICAL DATA SHEET #755 REV. 3

27120 SW 95th Avenue • Wilsonville, OR 97070 • 800.547.0659

TRICHOMONAS MEDIAPRODUCTS:Tubed Media:

Trichomonas Media, Feinberg T6399Trichomonas Media, Diamonds Modifi ed T3670, T6650Trichomonas Media, Diamonds Broth T6658Trichomonas 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 effi cacy 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 modifi ed by the incorporation of 0.1% w/v of agar which leads to reduced oxygen tension and consequently a much more prolifi c growth of trichomonads.

Diamonds Broth was developed to study trichmonads in animals and humans. The addition of antimicrobials helps supress microorganisms that would otherwise fl 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 Modifi ed 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 Kupferberg6,7 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 fi ltered water.

(1) Trichomonas Media, Feinberg:Liver Digest .....................................................25.0 gSodium Chloride ...................................................6.5Dextrose ...............................................................5.0Agar ......................................................................1.5Horse Serum ...................................................... 80.0 mlPenicillin .................................................1,000,000.0 unitsStreptomycin .............................................500,000.0 Final pH 6.5 ± 0.2 at 25°C

(2) Trichomonas Media, Diamonds Broth:Pancreatic Digest of Casein ................................20.0 gYeast Extract .......................................................10.0Maltose ................................................................. 5.0Dipotassium Phosphate ....................................... 0.8Monopotassium Phosphate .................................. 0.8Agar ...................................................................... 0.5L-Cysteine HCl...................................................... 1.0L-Ascorbic Acid ..................................................... 0.2Gentamicin ..........................................................20.0Vancomycin .........................................................20.0Horse Serum .....................................................100.0 ml

Final pH 6.0 ± 0.2 at 25°C



(3) Trichomonas Media, Diamonds Modifi ed: Same as (2) with the addition of 10.0 mg of Amphotericin B

(4) Trichomonas Media, Kupferberg:Pancreatic Digest of Casein ................................10.0 gPeptic Digest of Animal tissue .............................10.0Maltose ................................................................. 1.0Agar ...................................................................... 1.0L-Cysteine HCl...................................................... 1.5Chloroamphenicol .................................................0.1Methylene Blue .................................................0.003

Final pH 6.0 ± 0.2 at 25°C

PRECAUTIONS:*For in vitro diagnostic use. Observe approved biohazard precautions.

Storage: Upon receipt store at 2-8°C 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 specifi c 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 35-37°C 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 35-37°C.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 magnifi cation 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, Modifi ed and Kupferberg Trichomonas vaginalis (30001) Growth GrowthCandida albicans (10231) Growth Inhibition, partial to completeStaphylococcus aureus (25923) Inhibition, partial to complete Inhibition, partial to complete Diamonds Broth Trichomonas vaginalis (30001) GrowthEscherichia coli (25922) Inhibition, partial to complete Key: See “Interpretation”



User Quality Control: Check for signs of contamination and deterioration. Feinberg media should appear hazy and brown in color. Diamonds Broth and Diamonds Modifi ed 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.

bioMérieux, Inc.Data #755Revision Date: April 2009

trichomonas media

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