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Properties of Pseudomonas aeruginosa resistant to quaternary ammonium compound

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Title Properties of Pseudomonas aeruginosa resistant to quaternary ammonium compound
Names Washam, Clinton Jay (creator)
Sandine, William E. (advisor)
Date Issued 1968-03-14 (iso8601)
Note Graduation date: 1968
Abstract Pseudomonas aeruginosa cells were selected for their ability
to grow in the presence of 750 ppm alkyldimethylethylbenzyl
ammonium chloride (QAC). These cells were found to retain their
resistance to the germicide throughout tri-weekly transfers for 7
months in tryptone glucose yeast extract (TGY) broth containing no
QAC. Comparisons of the resistant and sensitive cells were made in
an attempt to define the mechanism of resistance and, in addition, to
provide some information as to the mode of action of QAC.
The germicidal activity of QAC solutions against both sensitive
and resistant cells in TGY broth was shown to be greatly affected by
the concentration of tryptone and yeast extract, but not by the amount
of sugar. The pH of the broth also influenced the germicidal activity;
both strains were more susceptible under slightly .
acid conditions. A
comparison of the pH range of growth of sensitive and resistant cells
demonstrated the ability of the former to grow in TGY broth at pH 4.5
while the latter could not achieve growth at pH 5.0. A study of the
effects of 50 ppm QAC, buffered to various pH levels, indicated that
the susceptibility of resistant cells was nearly the same as that of
sensitive cells below pH 3.0, at pH 6.0 and above pH 9.0. The
greatest difference between the two cell types occurred from pH 3.5
to pH 4.5 with a second peak of resistance being observed from pH
7.0 to pH 8.5. Electron microscopy revealed many dense inclusion bodies in
the resistant cells, some being near 0.2 μ in diameter. Furthermore,
the resistant cells were found to be much smaller (0.35 X 1.0 μ) than
the sensitive cells (0.75 X 3.0 μ). Sensitive cells possessed single
polar flagella while resistant cells were completely devoid of
flagella. Micrographs of sensitive cells exposed to 100 ppm QAC
exhibited no visible signs of lysis and the flagella did not appear to
be disrupted. In contrast, 10,000 ppm QAC caused abrupt lysis of
sensitive cells. A concentration of 1,000 ppm QAC had no visible
effect on resistant cells.
Broth cultures of the resistant strain displayed a distinct fruity
odor. Gas chromatographic analysis showed the QAC-resistant cells,
unlike the sensitive, produced large quantities of ethyl acetate and
ethyl valerate. Gel electrophoresis of cell-free extracts revealed
a difference in total protein and esterase patterns between the two
cell types. Two bands of esterase activity were demonstrated in sensitive cell extracts while only one band was detected in the
resistant cell extracts when alpha napthyl acetate was used as the
substrate.
Biochemical tests disclosed numerous differences between the
two cell types, many of which appeared to be interrelated. The most
significant differences were the losses in the ability of resistant cells
to synthesize extra-cellular lipase and protease enzymes. Many other
biochemical tests on resistant cells were negative or became positive
only after prolonged incubation. Permeability studies indicated a
greatly reduced rate of glucose uptake by resistant cells. Furthermore,
growth curve studies indicated a slower rate of growth by
resistant cells and a 15 minute longer generation time.
Genre Thesis/Dissertation
Topic Pseudomonas aeruginosa
Identifier http://hdl.handle.net/1957/46715

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