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A comparative study of Clostridium botulinum strains using differential egg yolk media

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Title A comparative study of Clostridium botulinum strains using differential egg yolk media
Names Stiebrs, Anita (creator)
Anderson, A. W. (advisor)
Date Issued 1967-05-03 (iso8601)
Note Graduation date: 1967
Abstract The purpose of this study was to compare and test the efficiency of the leading kinds of media being used for the isolation and identification
of Clostridium botulinum type E strains, and to develop a
more reliable method and media for the differentiation of this organism
based on the "pearly layer" characteristic.
The production of "pearly layer" around the colonies of the
different types of C. botulinum on egg yolk enriched media has been
used as an aid in differentiation and isolation of this group of bacteria. In this investigation a comparative study was conducted to test
the efficiency of the "pearly layer" formation on liver veal agar egg
yolk media (LVA) and trypticase agar egg yolk media (TA). Liver
veal agar egg yolk media was not entirely satisfactory in that strains
of C. botulinum type E and most non-toxic "type E-like" clostridia
did not always give a typical reaction. These organisms in many cases produced a faint
"pearly layer" or, in some cases, no "pearly layer" on the LVA egg yolk media. When trypticase was substituted as the base in the egg yolk media, more consistant results
were obtained in that all C. botulinum types produced the "pearly
layer". To separate type E strains from types A and B, sucrose and
an indicator, neutral red, were incorporated in the TA media. Since
only type E strains ferment sucrose, the acid in the media was detected
by the neutral red indicator as formation of a pink halo around
the colony. Unfortunately, the fermentation of sucrose in the media
suppressed the production of the "pearly Iayer". However, type E's
can be differentiated from other types on the sucrose TA egg yolk
media by the production of starch-Iike compounds which give an iodine
reaction. Types A and B and most non-toxic "type-E-like" clostridia
do not produce starch-like granules even on a utilizable sugar such
as glucose.
Trypticase base, therefore, is much more reliable than liver
veal base, not only "pearly layer" formation but also in differentiation
of C. botulinum type E's from other types by sucrose fermentation
and starch reaction.
Throughout this investigation, two morphologically different colony types have been associated with a pure culture of C. botulinum type E. One was the characteristic C. botulinum type E toxic colony which produced "pearly layer" on TA egg yolk plates; the other colony was a non-toxic sporulating isolate which did not produce
a "pearly layer". The toxic cultures produced acid and gas
from glucose, fructose , maltose, sucrose, and sorbitol. The nontoxic
isolates produced acid but no gas in the same sugars. Microscopically,
biochemically and morphologically the two types of
colonies were very similar to the toxic (TOX) cultures and the nontoxic
sporulating (OS) variant described by other investigators.
Genre Thesis/Dissertation
Topic Botulism
Identifier http://hdl.handle.net/1957/47433

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