Organization and Practice in Tuberculosis Bacteriology by C. H. Collins, J. M. Grange, M. D. Yates

By C. H. Collins, J. M. Grange, M. D. Yates

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Extra info for Organization and Practice in Tuberculosis Bacteriology

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48 Cultural methods Milk At least 100 ml of milk are centrifuged. The cream layer and the deposit are each treated by the NaOH or oxalic acid methods, centrifuged and the deposit cultured. Water Two litres of water are passed through a membrane filter. The filter is cut into strips not more than 1 cm wide and each strip placed in 3 per cent oxalic acid in a Petri dish for 15 minutes. They are then removed and placed in distilled water in another dish for 5 minutes. Each strip is placed on culture medium in a screw-capped bottle.

The number of other organisms is assessed either by examining a Gram-stained smear of the deposit or by culturing it on blood agar and incubating overnight. The deposit, meanwhile, is refrigerated. The deposit is suspended in 2 ml of sulphuric acid, 4 per cent. After standing for 15 to 40 minutes, depending on the number of non-acid fast organisms present, it is diluted with 15 ml of sterile distilled water and centrifuged again. The deposit is cultured. Pleural, cerebrospinal and other fluids The numbers of non acid-fast bacilli are assessed as for urines.

Pathological material that is known to contain other organisms must be treated in an attempt to destroy them but to preserve the mycobacteria. The reagents used must also reduce the specific gravity of the specimen so that it can be centrifuged with a reasonable certainty that the majority of the mycobacteria will be deposited. Unfortunately there are no reagents that will completely fulfil the first requirement and tuberculosis bacteriologists have learned to steer a middle course and to accept that if enough mycobacteria to give a diagnosis are to survive then a proportion of cultures will be contaminated with 'resistant' organisms such as Pseudomonas aeruginosa, some enterobacteria and streptococci and a range of yeasts and microfungi.

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