Artículo

J. clin. Path. (1957), 10, 327.

A CULTURE MEDIUM FOR TRICHOMONAS VAGINALIS DONNE AND SPECIES OF CANDIDA
BY

J. G. FEINBERG*

AND

M. JOAN WHITTINGTON

From the Lister Institute of Preventive Medicine, London, and the London Hospital, Whitechapel, London

(RECElVED

FOR PUBLICATION NOVEMBER

28, 1956)

The laboratory methods usually employed for detecting Trichomonas vaginalisin secretions are the microscopic examination of wet films or fixed, stained smears, and the culture of the parasites in artificial media. In recent years the cultural method has become more widely used as an aid in the diagnosis of trichomoniasis. Different culture media have been employed for this purpose with varying degrees of success, and this paper describes a medium which seems particularlysuitable, because it is cheap, easy to prepare and use, and keeps well. During the past 40 years many attempts have been made to grow T. vaginalis in a variety of media. At first simple liquid bacteriological media were used (Lynch, 1915; Reuling, 1921; Fischer, 1935, for example). Richer media of the type used by Boeck and Drbohlav (1925) have also given good results (Andrews, 1929; Hawes, 1947;Whittington, 1951, for example). All these media had the disadvantage of favouring the growth of the accompanying bacteria as well as that of the trichomonads. Johnson and Trussell (1943) and Sprince and Kupferberg (1947) used elaborate media for maintaining bacteria-free cultures of T. vaginalis. The value of these media for diagnostic purposes is unknown, however, and they are too complex andexpensive for ordinary routine work. McEntegart (1952) simplified Johnson and Trussell's (1943) formula for use in his studies on T. vaginalis, and Feinberg (1953) substantially modified McEntegart's medium and succeeded in growing the parasites in large numbers for experimental work. A further modification of this medium by Feinberg was suggested for use in routine diagnostic work, and the presentinvestigation has been carried out to assess its value.
*During the tenure of a U.S. Public Health Service Research Fellowship of the National Microbiological Institute.

Culture Medium The formula of this medium is:
Proteolysed livert Sodium chloride
Dextrose Inactivated horse serum Distilled water Penicillin . Streptomycin .

25.0 g. 6.5 g. 5.0 g. 80.0 ml. 1,000.0 ml. 1,000,000 units500,000 units

The solid components are dissolved in the distilled water, the serum added, and the pH adjusted to 6.4 by adding N/l sodium hydroxide (about 9 ml. per litre of medium). The mixture is sterilized by Seitz filtration and stored in screw-capped bottles in a refrigerator. In a diagnostic medium some method of suppressing the bacteria, which would otherwise overgrow the trichomonads, isdesirable. Antibiotics serve this purpose admirably, 1,000 units of penicillin and 500 units of streptomycin per ml. of medium being an effective concentration. No deterioration of the antibacterial powers of this culture medium is evident after three months' storage in a refrigerator at +40 to

+5° C.

Sources of Clinical Material
The patients from whom secretions were obtained were men andwomen attending the Venereal Diseases Department of the London Hospital (the Whitechapel Clinic), and women attending the Gynaecological Out-patient Department of the same hospital from June, 1954, to July, 1956. Samples of vaginal secretions were collected in small sterile plastic spoons (Jackson, Malleson, Stallworthy, and Walker, 1948). After a fresh smear had been made and examinedmicroscopically, the remainder of the sample was inoculated into about 7 ml. of the warmed culture medium in a 6 x i in. test tube. Urethral specimens from the men were obtained by Lanceley's (1954) method of gently rubbing the surface of the urethra with a platinum loop. Part
t" Panmede " brand, in powder form, obtained from Messrs. Paines and Byrne, Ltd., Pabyrn Laboratories, Greenford, Middlesex....