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Title: 牛乳培地による腟トリコモナスの分離培養について
Other Titles: On the Isolation Culture of Trichomonas vaginalis with Milk Media
Authors: 中林, 敏夫 / 宮田, 彬
Authors (alternative): Nakabayashi, Toshio / Miyata, Akira
Issue Date: Oct-1967
Publisher: 長崎大学熱帯医学研究所 / Institute of Tropical Medicine, Nagasaki University
Citation: 熱帯医学 Tropical medicine 9(2). p79-88, 1967
Abstract: Numbers of serum-free media have been so far devised for the isolation culture of Trichomonas vaginalis (Tv) and employed generally for the diagnosis of human trichomoniasis. These media, however, still have such an inconvenience that the aseptic harvest and storage of animal serum may be difficult under certain circumstances. An attempt was made of the isolation culture of Tv from vaginal secretion of patients with use of milk media, which contained cow milk but no serum, in order to examine on their capability of supporting the growth of this protozoa in comparison with a serum-containing medium, V-bouillon. Three media provided by the authors were derived from Samuels' C7 medium (1965) and called TTM, TCM and PCM in abbreviation. These were manufactured as described below: (1) TTM; 4.2g of trypticase, 1.2g of liver extract, 0.3g of yeast extract and 7.5mg of KH_2PO_4 were dissolved in 100ml of boiling distilled water. 8mg of citric acid, 20mg of thiomalic acid, 40mg of ascorbic acid and 0.5g of glucose were added to the solution after cooling. The solution was adjusted the pH to 6.0 with 1N NaOH and sterilized with a Seitz filter. It was further added with 0.8mg of cholesterol, which was dissolved in absolute ethanol at the rate of 20mg per ml, and 0.25ml of sterile milk, and distributed into sterile test tubes 5 ml per tube. (2) TCM; part A. 4g of trypticase, 1.2g of liver extract, 0.3g of yeast extract, and 0.6g of NaCl were dissolved in 80ml of boiling distilled water and autoclaved after the pH was adjusted to 6.0. Part B. 0.5g of glucose, 100mg of cysteine HCl and 40mg of ascorbic acid were dissolved in 20ml of distilled water and sterilized with a Seitz filter after the pH was adjusted to 6.0. Then, 0.8mg of cholesterol and 0.25ml of sterile milk were added to the solution. Both solutions of part A and B were mixed aseptically and distributed into test tubes. (3) PCM; this was quite similar in constitution as TCM with only exception that 4g of peptone was used instead of trypticase. In this experiment, a milk-free medium (TC), in which milk was omitted from TCM was used as a control. V-bouillon was a kind of modified CPLM medium and almost similar in constitution as CPLM, which was reported by Johnson, Trussell and John in 1945, and shortened as VB in this paper. The outline of results obtained was summarized as follows: 1) The detection rate of Tv in the isolation culture from 74 patients was equally 24.3% (18/74) with any of three milk media and was 25.7% (19/74) with VB. From this result, no significant difference in detection rate was recognizable between any of milk media and VB. 2) In the isolation culture of Tv with the milk media and VB, 80 to 90% of positive cases were confirmed within 48 hours of culture and the growth of Tv in the media was kept in satisfactory condition more than 96 hours of culture. 3) Some of the strains newly isolated from patients were subjected to an examination on the serial cultivation. The successive cultivation until the 8th generation was easily attained with use of TTM and VB, and the fairly good growth of Tv was usually demonstrated in both media. It was also possible with TCM and PCM in most of cases but these media could not be regarded as an appropriate medium for maintaining Tv, since the growth of Tv was usually lack of stability in the media. 4) The milk free TC medium appeared to demonstrate no good result in the examination on the isolation culture and serial cultivation as well. From the results described above, it was concluded that an indispensable component for the growth of Tv might be contained in milk and TTM medium would be practically usable either in the isolation culture or in the serial cultivation of Tv.
URI: http://hdl.handle.net/10069/4012
ISSN: 03855643
Type: Departmental Bulletin Paper
Appears in Collections:Volume 9, No. 2

Citable URI : http://hdl.handle.net/10069/4012

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