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Antimicrobial Agents and Chemotherapy, March 1999, p. 655-660, Vol. 43, No. 3
Departments of Microbiology & Molecular
Genetics and Medicine, University of California Irvine, Irvine,
California 92697-4025
Received 28 May 1998/Returned for modification 18 September
1998/Accepted 13 December 1998
The spirochete Borrelia burgdorferi was unexpectedly
found to be as susceptible to diacetyl chloramphenicol, the product of the enzyme chloramphenicol acetyltransferase, as it was to
chloramphenicol itself. The susceptibilities of Escherichia
coli and Bacillus subtilis, as well as that of
B. burgdorferi, to diacetyl chloramphenicol were then
assayed in different media. All three species were susceptible to
diacetyl chloramphenicol when growth media were supplemented with
rabbit serum or, to a lesser extent, human serum. Susceptibility of
E. coli and B. subtilis to diacetyl
chloramphenicol was not observed in the absence of serum, when horse
serum was used, or when the rabbit or human serum was heated first. In
the presence of 10% rabbit serum, a strain of E. coli
bearing the chloramphenicol acetyltransferase (cat) gene
had a fourfold-lower resistance to chloramphenicol than in the absence
of serum. A plate bioassay for chloramphenicol activity showed the
conversion by rabbit, mouse, and human sera but not bacterial cell
extracts or heated serum of diacetyl chloramphenicol to an inhibitory
compound. Deacetylation of acetyl chloramphenicol by serum components
was demonstrated by using fluorescent substrates and thin-layer
chromatography. These studies indicate that esterases of serum can
convert diacetyl chloramphenicol back to an active antibiotic, and
thus, in vitro findings may not accurately reflect the level of
chloramphenicol resistance by cat-bearing bacteria in vivo.
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Esterases in Serum-Containing Growth Media
Counteract Chloramphenicol Acetyltransferase Activity In
Vitro
and
*
Corresponding author. Mailing address: Departments of
Microbiology and Molecular Genetics and Medicine, University of
California Irvine, Irvine, CA 92697-4025. Phone: (949) 824-5626. Fax:
(949) 824-8598. E-mail: abarbour{at}uci.edu.
Present address: Tuberculosis Research Laboratory (151), Veterans
Administration Medical Center, Long Beach, CA 90822.
Antimicrobial Agents and Chemotherapy, March 1999, p. 655-660, Vol. 43, No. 3
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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