緑膿菌のピオシン型別に関する研究-4-型別法2種を併用しての培地の検討および集落型の差によるピオシン産生性と感受性
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Both the methods of Darrell-Wahba and Gillies-Govan were used in parallel for pyocine typing of test-strains in the present study, consisting of 8 indicators (G1-G8) and 2 type strains (Ty.1 and Ty.16) for the G-G method and of 12 indicators (W1-W12) for the D-W method. In the G-G method defibrinated horse blood was used throughout the present study. The first experiment was carried out to compare those activities of pyocines produced, with one another, on slightly modified media as described in the remarks for Tables 2 and 4, using the similar slant culture originatad from a la colony of each test-strain. No different patterns were observed on strains shown in Tables 1 and 3. In tables 2 and 4, the first pairs of inhibition patterns for each strain were obtained by the first typing of the experiment, the second pairs were observed by the typing on 2 days later, and the third ones were showed by the typing performed 5 days after the second typing. From these results, it can be said that trypto-soy agar (Eiken) was usefull in place of tryptone soya agar (Oxoid) for both the typing methods and horse blood stored in the refrigerator for 3 weeks had no trouble with the use for the G-G method. It may be worth to pay regard, however, to the result showing that there were some strains producing different pyocines after the storage for one week. Basing upon the reasons obtained from the experiment and after due consideration of the results, trypto-soy agar was used in the further experiments. Secondly, 6 slants among the test-strains were subjected preliminarily to the comparison of pyocine productivities with one another on the media different from the original method 2 days after the third typing of the first experiment. Excepting G1 from Table 5 and Ty.16 from Table 6, the same pattern in each strain was observed on a different medium each, but many more strains should be to be examined before the definite conclusion can be safely drawn. The pyocine produced by la and sm colonies and ordinary subcultures of 11 teststrains were studied and typed using 3 sets of indicator under the same cultural condition as the producers on the day. Inhibition patterns observed by different colonial conditions of the producers (Tables 7-10) and sensitivities to the pyocines of the 3 sets of indicator (Tables 11-14) were fairly complicated. Differences of the inhibition patterns between la and sm types of producers were observed in 9 cases by the G-G method (Table 8) and 14 cases by the G-W method (Table 10), and different sensitivities between la and sm sets of indicator were seen in 13 cases by former method (Table 12) and in 19 cases by later method (Table 14). These findings indicate that the colonial type of indicators may be regard as more weighty factor to obtain reproducible results than that of the producers. At any rate, fresh la colonies should be selected and used for the pyocine typing, irrespective of being test-strains to be typed and or of being indicator strains. In the fourth experiment, a la colony of each indicator and 9 la colonies of type strains were typed using 2 sets of indicator originated from different la colonies selected just before the use. According to the G-G method (Table 15), 33 out of 38 colonies showed no difference between the patterns obtained by 2 sets of indicator, whereas by the D-W mehtod (Table 16), different patterns were observed among 12 out of 29 colonies excepting the results of Ty. 16. From the observation above, it can safely be said that no difference in sensitivity to pyocine and production of pyocine were observed among different la colonies.
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- 1972-06-00
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