The influence of cultural conditions on the production of diphtheria toxin / A.F. Watson and Elsie Langstaff.

  • Watson, A. F.
Date:
[1927?]
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Licence: In copyright

Credit: The influence of cultural conditions on the production of diphtheria toxin / A.F. Watson and Elsie Langstaff. Source: Wellcome Collection.

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    1 II P. /f-S I Reprinted from The Journal OF PATHOLOGY AND BACTERIOLOGY, Vol. XXX., 1927 j THE INFLUENCE OF CULTURAL CONDITIONS ON THE PRODUCTION OF DIPHTHERIA TOXIN. A. F. Watson and Elsie Langstafe. The Wellcome Phijsiological Research Laboratories, Beckenham, Kent. I I.—The development of cultueal methods foe the eoutine PEEPAEATION OF DIPHTHEEIA TOXIN AND TOXOID. A few years ago Hartley and Hartley (1922) described a general ‘ method for the preparation of diphtheria toxin and its results in their hands. They emphasised the importance of the production of satisfactory antigens in laboratories where the preparation of thera¬ peutic sera is carried out on a large scale. Since these results were published certain advances in this field have been made and a general survey of the work is included in the three parts of this paper. The present part deals especially with the development of methods although, in devising these, advantage has been taken of work described in the second and third papers. These methods were made possible by the use of the Eamon (1922) flocculation test for titrating > culture filtrates of C. diphtherice against antitoxin in vitro. Over a long period individual culture filtrates were tested and two aspects of importance in routine production were soon evident. It was found that the variation from filtrate to filtrate from the same medium autoclaved atjoH = 8'0 (especially with media made by tryptic digestion of muscle) might be very great {infra, p. 394). In attempting to trace the cause of this variation, the importance of the heat stability of media became prominent. Although the ill effects of heat on media have been reported by many workers, the influence of autoclaving at joH = 8’0 in contributing to this variation had not been suspected by < us {infra, p. 405). It was decided at the beginning of the work to carry out a detailed investigation of the construction of medium by the tryptic digestion : of muscle, a process which Hartley (1922) adapted to the preparation : of diphtheria toxin from Douglas5 (1914) original method. The media used in this work and that in the following two papers were prepared by two modifications of enzyme digestion and by dissolving 2 per cent, of commercial peptone X in beef-heart infusion.* * The beef-heart infusion was prepared by boiling muscle and water in the proportion of 1 lb. of minced muscle to 1 litre of water. The infusion was filtered, | 2 per cent, peptone and 0*5 per cent, salt added and the pH adjusted to 8-0. After filtering the medium, sterilisation, method of inoculating etc. were carried out as for digest media.
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