抗酸菌の呈する硝酸塩還元反応

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Nitrate reduction activity was tested on thestock cultures of various Mycobacteria. For thequantitative measurement of nitrite, three reagents were used following Shinns method: one to two dilution of HC1, 0.2% sulfanilamide solution and 0.1% N (1-Naphthyl) ethylendiaminedihydrochloride solution. As NaNO2 solutionshows a lineality between the concentrations from10-6M to 6×10-6M, the reaction was read quan titatively by the use of photometer. All strains tested were subcultured on 1% Ogawas medium and the growth was used forthe test. One to hundred mole solution of NaNO3 in 1/40 M phosphate buffer (pH 7.0) was autoclaved at 120°C for 20 minutes and stored atroom temperature. Thirty ml of NaNO3 solutionthus prepared was put into a test tube and 60mg of bacillary mass of each strains was addedaseptically into it. After every 2, 4, 6 and 24 hoursof incubation at 37°C, the amount of NaNO2produced was measured quantitatively. Stock cultures tested were as follows: 11strains of Mycobacterium tuberculosis including8 drug resistant strains, 3 strains of M. bovis, 8 substrains of BCG, 8 strains of M. avium, 6saprophytic and 17 unclassified acid-fast bacilli.Among these unclassified Mycobacteria, 5 werePhotochromogens, 6 Scotochromogens, 6 Nonphotochromogens according to Runyons Classification. In accordance with their type and group, theseMycobacteria have shown various degrees of activity in reducing nitrate. Both drug sensitive and drug resistant strainsof M. tuberculosis have shown strong activity. Contrary to this, M. bovis could not show theactivity in reducing nitrate after 6 hours of incubation. Among 8 BCG strains, however, somehave shown strong and others weak activities inreducing nitrate. Among 8 strains of M. avium, 7 strains couldnot reduce nitrate at all even after 24 hours ofincubation, while only one (Flamingo) has shownweak positive reaction. Den-Cho strain, which was originally isolatedfrom chicken and maintained for a long periodof time as an avian strain, has to be identifiedas a saprophytic strain not only with its strongactivity in reducing nitrate but also with otherbiological properties. The strong positive reaction was observed inthe strains of M. phlei and M. smegmatis. M.fortuitum was also able to reduce nitrate relatively strong, but M. ranae was able to do soonly weakly. Among the unclassified Mycobacteria, Photochromogenic strains alone have shown strongactivity, whereas both Scotochromogenic and Nonphotochromogenic strains very weak activity. From these results, it is concluded that nitratereduction test seems to be available in the dailylaboratory testing as one of the methods in differentiating qualitatevely the type or the groupof Mycobacteria to be examined.

著者

佐藤直行
国立予防衛生研究所

抗酸菌の呈する硝酸塩還元反応

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