腟トリコモナスの培養における牛乳培地の検討

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In a majority of culture media hitherto devised for Trichomonas vaginalis (Tv), use of animal sera was inevitable to meet the nutritional requirement of this parasite. In this meaning, Samuels'C_6 and C_7 medium (Samuels, 1963, 1965), in which light cream or whipping cream was used for serum, was of much interest. This experiment was done to examine Samuels'C_7 and further to make up an improved milk medium. A serum-containing medium (V-Bouillon) was used in this experiment as a control. The outline of results obtained was summarized as follows: 1) Subcultures of Tv with modified C_7 media in which various milk products were used in place of whipping cream were found impossible. It was, however, observed that fresh cow milk fairly stimulated the growth of Tv (Table 1). 2) As an improved C_7, TTM medium (Trypticase-Thiomalic acid-Milk Medium) which was finally prepared through the quantitative examination of each medium component demonstrated an excellent capability of supporting the growth of Tv. TTM medium was prepared as described below: 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 1 N NaOH and sterilized with a Seitz filter. To the solution was further added 0.8mg of cholesterol, which was dissolved in absolute ethanol at the rate of 20mg per ml, and 0.25ml of fresh milk which was sterilized by autoclaving at 120℃ for 20min. or by intermittent boiling at 100℃ for 30min. 3 times, and distributed into sterile test tube　5ml per tube. (Tables 2 and 3). 3) The following substances were found to be essential for Tv cultivation in TTM: cow milk, cholesterol, ascorbic acid and thiomalic acid. From the fact that thiomalic acid could be replaced by cysteine HCI, it was assumed that these substances might play a role in lowering the oxidation-reduction potential of medium. (Tables 3, 4, 5, 6 and 7). 4) It was thought that trypticase and liver extract were the important sources of amino acids in TTM. Trypticase could not be completely replaced by polypeptone but by each 2g of trypticase and polypeptone, the growth of Tv was satisfactorily promoted. (Tables 8, 9 and 10) From the results described above, it was concluded that TTM medium would be practically useable in the serial cultivation of Tv. The principal differences of TTM from C_7 medium were in; firstly the sterilization of medium with the Seitz filter apparatus, secondarily the addition of cow milk instead of whipping cream and lastly adjusting the pH to 6.0. As to isolation culture of Tv from vaginal secretion of patients with use of TTM and other two milk media derived from C_7 medium, a report was previously published by the present authors (Nakabayashi and Miyata, 1967).
長崎大学熱帯医学研究所,Institute of Tropical Medicine, Nagasaki Universityの論文
1968-03-30

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中村敏夫
長大・熱研・疫学

腟トリコモナスの培養における牛乳培地の検討

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