DIAMOND BROTH

Technical Data #2351a / 98.06.25

Diamond Broth is selective media used for the detection and cultivation of Trichomonas sp. Inhibitors suppress bacterial growth and permits cultivation of Trichomonas. Amphotericin B inhibits the growth of Candida sp.

FORMULA
in grams per liter purified filtered water

Casein Peptone	20,0
Yeast Extract	10,0
Potassium Phosphate, dibasic	0,8
Potassium Phosphate, monobasic	0,8
L-Cysteine HCl	1,0
Ascorbic Acid	0,2
Maltose	5,0
Agar	0,5
Horse Serum	100,0 ml
Antibiotics
Vancomycin	0,02
Gentamycin	0,02

pH 6,0 +/- 0,2 à 25º C

This approximate formula may be adjusted and/or enriched to obtain best results.

PRECAUTIONS
This medium is for IN VITRO diagnostic use only.

STORAGE
Store prepared media at 2-8º C protected from direct light and dehydrated powder, in a dry place in tightly-sealed containers at 2-25º C.

SIGN OF DETERIORATION
Media should not be used if the expiry date has passed. Prepared media should not be used if there are signs of contamination (evaporation or discoloration). Do not use dehydrated media if it is caked.

DIRECTIONS
Suspend 38,3 g in 900 ml of purified filtered water. Heat with frequent agitation and boil for one minute. Sterilize at 121º C for 15 minutes. Cool to 45-50º C. Add 100 ml of Horse serum (4271) and sterile antibiotics (8713). Mix thouroughly and dispense into sterile culture tubes.

PROCEDURE
Bring prepared medium at room temperature before inoculation.

1. Directly, inoculate with specimens from vaginal, urethral and prostatic secretions or with sediment from centrifuged urine.
2. Incubate at 35º C for 24-72 hours.
3. Examine daily microscopically. One drop on a slide covered with coverslide should be examined at low magnification.
4. Discard after 3 days if negative.

QUALITY CONTROL
Expected results after 24 hrs at 35ºC

Organisms	ATCC	Growth
Trichomonas vaginalis	clinical	+
Escherichia coli	25922	- or partial

LIMITATIONS OF METHOD
This medium is only a part of the identification. Other tests may be required.

REFERENCES

1. Bowie, W.R., G.E. Garber, L. Sibau, R. Ma, E. M. Proctor, C.E. Shaw 1987 Cell culture compared with broth detection of Trichomonas vaginalis J. Clin. Microbiol. 25:1275-1279
2. Lennette, E.H., Ballows, A., Hausler, W.J.Jr., and Shadomy, H.J. Manual of Clinical Microbiology. 4th ed. 1985 Washington D.C.: American society for Microbiology.
3. Rashad, A.L., 1989 Conference on antimicrobial agents and chemotherapy, Abstracts of the annual meeting. No. 517, Houston, p.187
4. Schmid, G.P., L.C. Matheny, A.A. Zaidi, S.J. Kraus 1989 Evaluation of six media for the growth of Trichomonas vaginalis from vaginal secretions J. Clin. Microbiol. 27:1230-1233
5. Thomason, J.L., S.M. Gelbart, J.F. Sobun, M.B. Schulen, P.R. Hamilton 1988 Comparaison of four methods to detect Trichomonas vaginalis 26:1869-1870
6. Thomason, J.L., S.M. Gelbart, P.J. Osypowski, J.A. James, P.R. Hamilton 1989 Comparaison of Diamond's medium modified and Kupferberg medium for detection of Trichomonas vaginalis 27:1095-1096

CATALOGUE NUMBERS

Dehydrated media	500 g	QB-39-1114
Horse Serum	100 ml	4271
Diamond supplement, 5ml	10/pkg	8713
Prepared media
Tubes 16x125 mm, 5 ml	10/pkg	2351

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