The Failure of Bacterium coli from Human Feces to Grow at 46° in the Eijkman or the Bulír Tests

Abstract

The Eijkman test (1904) is designed to detect fecal pollution of water. Water is inoculated into glucose-peptone broth which is incubated at 460, and gas production is assumed to indicate fecal pollution from warm blooded animals. Bacterium coli and Bacterium aerogenes strains of non-fecal origin or from feces of coldblooded animals are not supposed to grow, or at least not to produce gas under these conditions. Although much has been written in its favor and much against the test, it would seem that the fundamental question as to whether or not most of the colon bacilli from human feces are able to grow and produce gas at a temperature of 460 has not been satisfactorily answered. A review of both sides of the question will be found in the papers of Leiter (1929), of de Magalhaes (1932), and of the authors (1930). The latter, by using the dilution method, showed that in many samples of feces, at least half of the bacteria capable of producing gas from lactose at 37.5°refused to produce any at all from glucose at 460, and that in some samples of feces very few, even less than 1 per cent of the fecal B. coli, were able to produce gas under the conditions of the Eijkman technique. The Bulir test (1907) is a modification of the Eijkman test, and consists in using a mannitol-peptone beef infusion broth at 460.

Decomposition of Carbohydrates and Alcohols with Production of Gas at 46°C. By Members of the Genus Escherichia

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Although it has been known for many years that Escherichia coli grows rapidly and luxuriantly at a temperature well above that of the body, only a few workers (Leiter, 1929;Williams, Weaver, and Scherago, 1933;Hajna, 1933 andHajna and Perry, 1935) have utilized higher temperatures either in the isolation of this organism or in the study of its fermentation of various carbohydrates or alcohols. As far as the author is aware, only glucose (Eijkman, 1904;Hajna, 1933, Hajna andSkinner and Brown, 1934), mannitol (Bulir, 1907;Minkewitsch, 1929;Skinner and Brown, 1934), and lactose (Levine, Epstein and Vaughn, 1934) have been utilized in the investigation of the ability of Escherichia coli to ferment at, or around, the temperature of 460C.The following investigation was therefore undertaken to determine whether the members of the genus Escherichia can decompose carbohydrates and alcohols other than those referred to, at this temperature.

METHODS AND MATERIALSCultures. Cultures isolated from human feces, were grouped on the basis of (1) fermentation of sucrose, sorbitol, dulcitol, adonitol, and salicin at 370C.

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“…After the method ofSkinner and Brown (1934). t Basic medium ofHajna (1933 and.Note: Final pH of all media 7.0.…”

A Comparison of Standard Lactose Broth with Lauryl Sulphate Broth and with the Eijkman Method for Demonstrating Fecal Coliform Bacteria1

“…By fecal coliform bacteria we mean simply coliform bacteria from feces, not coliform bacteria that may have or have not, this or that biochemical property. It has been shown that the original Eijkman method allows only a small proportion of the fecal coliform bacteria to grow and produce gas (Skinner and Brown, 1934). At various times the test has been modified by changing the formula of the medium, and by altering the incubation temperature.…”

A Comparative Study of Standard Lactose Broth and Modified Eijkman Medium for Isolation of Escherichia Coli From Nut Meats

“…Subsequent studies by these investigators (1935) further indicated the superiority of the Eijkman medium over standard lactose broth in the examination of oysters, crabmeat, and other foods. They therefore suggested that in the examination of those products where Escherichia coli is the sole basis for estimating probable fecal pollution, the Eijkman medium be substituted f o r standard lactose broth.The limitations of the Eijkman test for the detection of Escherichia coli from human feces was shown by Skinner and Brown (1934). Two-thirds of the samples examined by them contained thermolabile strains of Escherichia coli which were recovered with standard lactose broth but which failed to survive the incubation temperature of the Eijkman method.Minkevich, Alexandrov, and Soboleva (1936), confirmed the findings of Skinner and Brown regarding the injurious influence on Escherichia coli of 46°C.…”

“…The limitations of the Eijkman test for the detection of Escherichia coli from human feces was shown by Skinner and Brown (1934). Two-thirds of the samples examined by them contained thermolabile strains of Escherichia coli which were recovered with standard lactose broth but which failed to survive the incubation temperature of the Eijkman method.…”