The bacterial diseases of respiration, and vaccines in their treatment / By R. W. Allen.

  • Allen, R. W., 1876-1921.
Date:
1913
Catalogue details

Licence: Public Domain Mark

Credit: The bacterial diseases of respiration, and vaccines in their treatment / By R. W. Allen. Source: Wellcome Collection.

Provider: This material has been provided by the Augustus C. Long Health Sciences Library at Columbia University and Columbia University Libraries/Information Services, through the Medical Heritage Library. The original may be consulted at the the Augustus C. Long Health Sciences Library at Columbia University and Columbia University.

    34/276 (page 20)
    has named splitter are well demonstrated; these frequently appear as minute red granules, and are probably infant bacilli. By it I have frequently been enabled to detect tubercle bacilli in sputum which other methods have entirely failed to reveal. The antifovmin method has been devised not only for the concen- tration of the tubercle bacilli in a specimen of sputum, so that very scanty numbers may be detected with ease, but also for the obtaining of pure cultures of the tubercle bacilli direct from the sputum; it per- forms both ends with conspicuous success. Antiformin is a mixture of equal parts of a 15 per cent, aqueous solution of caustic soda and the liquor sodse chlorinatas of the B.P. The procedure is as follows : 5 c.c. of the sputum are placed in a sterilised test-tube supplied with a well- fitting rubber cork; to this 5 per cent, of a 30 per cent, aqueous solution of antiformin are added, the tube is stoppered, well shaken till all the sputum is dissolved and a homogeneous mixture secured. It is then placed in the incubator at 37° C. for one hour. The liquid is distri- buted into two sterile centrifuge tubes and centrifuged at a high speed for five minutes, the supernatant liquid is pipetted off, to the sediment in each tube 5 c.c. sterilised distilled water are added, and thorough centrifugalisation performed; this procedure is twice repeated. The final sediment is employed for the insemination of tubes of Dorset's egg medium, and for the preparation of smears which are stained according to one or other of the preceding methods—best by Spengler No. II. Personally, I add 5 c.c. of the mixture after incubation to one centrifuge tube, and 2 or 3 c.c. only to the other, making the volume up to 5 c.c. with absolute alcohol. The latter tube has a distinctive mark, and subsequent washings of the sediment are carried out with 40 per cent, alcohol. The addition of the alcohol lowers the specific gravity of the liquid, and facilitates the deposit of the tubercle bacilli; this tube cannot be used for culture purposes, but only for the pre- paration of smears. The pepsin and trypsin methods depend upon the digestion of the sputum with acid pepsin or alkaline trypsin prior to centrifugalisation. In opposition to the view of their advocates I do not think they have any advantage whatever over the antiformin method, which, in my hands as well as those of others, has proved most reliable and valuable. Leishuuin's method has been already described on p. 13, and serves well to display the spirochastes, vibrios and spirilla of the mouth; ]f more intense staining be desired, a 10 per cent, aqueous solution of carbol-fuchsin may be applied for forty-five seconds, followed by thorough washing in running water. Even in this dilution carbol-fuchsin is both an intense and diffuse stain; it must therefore be remembered that the bacteria as seen under the microscope will appear unduly large. The