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In vitro identification of Clostridium botulinium by means of extracellular enzyme tests

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Title In vitro identification of Clostridium botulinium by means of extracellular enzyme tests
Names Levant, Emory Marion (creator)
Anderson, A. W. (advisor)
Date Issued 1969-05-07 (iso8601)
Note Graduation date: 1969
Abstract The purpose of this study was to examine the culture
supernatants of various strains of Clostridium botulinum
for characteristic extracellular enzymes which can facilitate
their rapid identification.
A convenient, efficient, water-cooled flat gel electrophoretic
unit was utilized to concentrate and purify the
total proteins and extracellular enzymes present in the
supernatants of various strains of C. botulinum types A, B,
C, E and F. The protein-laden gels, after staining, offer
evidence for characterization of the strains and possible
taxonomic significance.
Variations in characteristic proteolytic isozymes,
lipases, lecithinases, iron-bound proteins and diaphorases
reinforce the separation and identification of C. botulinum
types A, B, C, E and F. The variations in numbers and
electrophoretic mobilities of the proteolytic isozymes
suggest a usefulness in differentiating types A and B.
This detectable proteolytic property is used to separate
type A and type B organisms from the other known types of
C. botulinum. In addition, the detection of a single NADH
dependent nitroblue tetrazolium reductase band is indication
of a type A or B form. Two such diaphorase bands were
found in the supernatants of toxic type E and type F microorganisms.
Under the conditions of these investigations
the non-toxic forms did not show diaphorase activity.
Dehydrogenases characteristic of citric acid cycle metabolism
were not detectable in the supernatants using the
methods of these investigations.
Evidence accumulated using inhibitors and stimulants
suggest certain physiological roles for the diaphorases
found in the supernatants.
The NAD dependency of the diaphorases was established
following substitution of NADP into the assay systems.
Genre Thesis/Dissertation
Topic Clostridium botulinum
Identifier http://hdl.handle.net/1957/46071

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