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Quantitative determination of ristocetin by a microbiological diffusion method

Количественное определение ристоцетина диффузным микроБиологическим методом

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Summary

The basic factors influencing the microbiological diffusion titration of ristocetin were studied. The results were used as a basis for the elaboration of directions for the routine determination of this antibiotic in fermentation liquids, usingBacillus subtilis as the test microorganism and spontin as the standard. The basic conditions for obtaining sharply defined inhibition zones and reproducible results are as follows:

  1. (1)

    Titration medium without the addition of glucose; pH after sterilization 6.5;

  2. (2)

    Density of the inoculum corresponding to 3.2×107 to 5×107 spores/ml;

  3. (3)

    Preculture (60 min. at 20°C) of the titration medium, together with the specimens, so as to reduce the unfavourable effect of diffusion of the antibiotic on the quality of the inhibition zones;

  4. (4)

    Lowering of the sodium chloride, potassium chloride and copper sulphate concentration in the analysed specimens by diluting them to the maximum degree possible.

Abstract

Исследовались основные факторы, влияющие на микробиологическую диффузную титрацию ристоцетина. На основе полученных результатов была разработана инструкция для рутинных определений этого антибиотика в ферментационных средах с применением Bacillus subtilis в качестве тест-микроба и спонтина в качестве стандарта. Основными условиями для получения отчетливых зон ингибиции и воспроизводимых результатов являются:

  1. (1)

    Титрационная среда без прибавления глюкозы с pH 6,5 после стерилизации.

  2. (2)

    Густота используемго инокулума, отвечающая 3,2×107–5×107 спор в 1 мл.

  3. (3)

    Предварительная культивация (60 мин. при 20° C) титрационной панели с нанесенными на нее образцами с целью ограничения неблагоприятного влияния диффузии антибиотика на качество зон угнетения.

  4. (4)

    Понижение концентрации хлористого натрия, хлористого калия и сернокислой меди в анаизируемых образдах путем максимального возможного разведения образцоб.

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Authors and Affiliations

  1. Department of Microbiology, Institute of Biology, Czechoslovak Academy of Sciences, Prague 6

    Z. Řeháček

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  1. Z. Řeháček
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Řeháček, Z. Quantitative determination of ristocetin by a microbiological diffusion method. Folia Microbiol 6, 22–26 (1961). https://doi.org/10.1007/BF02869591

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  • DOI: https://doi.org/10.1007/BF02869591

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