The effect of temperature of incubation on the results of tests for differentiating species of coliform bacteria

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Abstract
1. A study has been made of different types of coliform bacteria with particular reference to (a) their ability to ferment lactose in MacConkey broth at different temperatures, and (b) the effect of using different temperatures of incubation for the indol, methyl-red, V.P., and citrate tests.2. It was found that 97% of the cultures ofBact. coli(indol positive, methyl-red positive, V.P. negative, citrate negative) examined could ferment lactose with production of acid and gas between 40 and 44° C. The number was not appreciably reduced at 45° C. but was markedly reduced at 46° C. 28% of the cultures ofBact. coli(indol negative, methyl-red positive, V.P. negative, citrate negative) and 15% ofBact. aerogenes(indol negative, methyl-red negative, V.P. positive, citrate positive) were found to be positive at 44° C.3. The adoption of a temperature of incubation of 30°C. for the V.P. test as advocated by Levine (1941) and the use of O'Meara's test showed that many cultures previously regarded as unable to produce acetylmethylcarbinol were in fact able to do so. Employing a temperature of 30°C. for 5, or in some cases 7, days for the methyl-red test, it was found that with nearly all the cultures tested there was an inverse correlation between the results of the methyl-red test and those of the V.P. test. With these modifications in technique some cultures originally designated as Intermediate type I were found to have reactions corresponding with those ofBact. aerogenestype I. Similarly, many cultures originally classified as Intermediate type II should have been typed asBact. aerogenestype II.4. It was found that all cultures of Intermediate type I classified as such by the new technique were incapable of using the nitrogen of uric acid for growth, but that the majority produced hydrogen sulphide. Cultures ofBact. aerogenestype II, on the other hand, grew well in uric acid medium, but produced no hydrogen sulphide.The investigation described in this paper was carried out by the Freshwater Biological Association, as part of the programme of the Water Pollution Research Board of the Department of Scientific and Industrial Research. The paper is published by permission of the Department.

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