Growth of Anaerobes in Crystal Violet Bile Media

Abstract

While experimenting with crystal violet bile media for use in water analysis, it was thought that some information could be obtained by testing the growth of anaerobes in media containing crystal violet and bile. An ideal medium for the detection of pollution in water supplies would be, of course, a medium which would prevent the growth of all bacteria except members of the colon group. This, however, is far from possible. B. welchii and other anaerobes are responsible for many false presumptive tests in waters. This fact was first reported by Frost (1906) in 1906. Many investigators have observed this condition in recent years. The question arose as to whether crystal violet bile broth would be effective in preventing the growth of anaerobes. Apparently this subject has not been investigated in regard to water analysis as no reference could be found in the literature. Hall and Ellefson (1918) (1919) were the first to call attention to the fact that gentian violet, when added to lactose broth, would eliminate the false presumptive test due to anaerobes. They found that one part in 20,000 was effective in eliminating all gas-forming anaerobes. Wagner and Monfort (1921) confirmed the work of Hall and Ellefson. EXPERIMENTAL Lactose broth was prepared and varying amounts of crystal violet and bile were added. The strength of the lactose and peptone was 1 per cent each. The pH value of all media was 117

Growth of Anaerobes in Bile Media Containing Malachite Green and Brilliant Green

“…A great amount of gas was produced in all of the media with a bile content greater than 1 percent even when the dye content was as great as 1: 1,000. The cultures of Type II were also found to be resistant to crystal violet as reported in a previous communication (2). The cultures of Type III and Type IV varied greatly.…”

“…He found, however, that the former strength afforded sufficient protection in most of the water supplies studied. One of us (2) while working with the crystal violet bile media found that a very high concentration of this dye was necessary to eliminate anaerobes. 'With a number of cultures of anaerobes a dye content greater than 1: 1000 was required to prevent growth and gas formation.…”

333. The coliform flora of milk and dairy products

“…Hall & Ellefson(2), and Salle(3) have proposed the addition of brilliant green, gentian violet, or crystal violet to the test medium as inhibitors, but Stark & England (4) found that this procedure suppressed some faecal coliform types. Poe (5) found that addition of bile salt to the medium did not eliminate anaerobes and sometimes encouraged their growth. However, Wilson (i), in an extensive, critical examination of the coliform test, states that bile salt-lactose broth yielded few false positives from water or milk, and this is probably the common experience in Britain.…”

“…Later Oxley(i9) showed that such strains occur commonly in raw milk. (5) The existence of 'heat-resistant' coliforms, though of no significance as regards actual testing methods, may be important in dairy-plant control, and recently this problem has attracted some attention. The heat resistance of coliform organisms in milk had been studied by, among others, Ayers & Johnson (20), Tanner & Windsor (21), Wilson(i), and Vernon & Walker (22); in ropy milk by Hammer & Hussong(23); and in ice cream by Nelson et al (24) and Paley & Isaacs (25).…”