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Biomedical subjects

D H Bouanchaud

Publications and source records attributed to D H Bouanchaud.

At least 37 records · Page 2Linked to original sources

Mechanism of transferable resistance to chloramphenicol in Haemophilus parainfluenzae.

A clinical isolate of Haemophilus parainfluenzae resistant to chloramphenicol and tetracycline transferred both cam and tet determinants to Escherichia coli K-12 during mixed cultivation on solid media irrespective of the selection employed. The doubly resistant transconjugant was found to contain levels of the enzyme chloramphenicol acetyltransferase (CAT) comparable to those found in R plasmid-bearing chloramphenicol-resistant enteric bacteria. Purification of CAT from the transconjugant was achieved by affinity chromatography, and the electrophoretically homogeneous protein was compared with previously characterized CAT variants specified by R plasmids. Although the CAT associated with cam from H. parainfluenzae was found to be distinct from the three types described previously, its N-terminal peptide amino acid sequence was identical with that determined for a type II CAT. Attempts to demonstrate covalently closed circular deoxyribonucleic acid in the H. parainfluenzae donor and the E. coli transconjugant were unsuccessful. The cam and tet determinants were nontransmissible from E. coli but could be cotransferred following the introduction of a suitable conjugative plasmid.

Cell-Free System↗

Chloramphenicol resistance in Streptococcus pneumoniae: enzymatic acetylation and possible plasmid linkage.

Clinical isolates of Streptococcus pneumoniae resistant to chloramphenicol were observed in France for the first time in 1973. During a 4-year survey, these strains were found to represent 6% of a total of 564 isolates of S. pneumoniae in a general hospital and to belong to 13 different serotypes. One such strain, referred to as BM 6001, was shown to inactivate chloramphenicol, and the process was found to be inducible. The inactivated products were demonstrated to be O-acetoxy esters of chloramphenicol. The synthesis of an inducible chloramphenicol acetyltransferase was shown to be responsible for the inactivation of the drug. The resistant strain was able to transfer the chloramphenicol marker by transformation to competent strains of pneumococci at a frequency of 1% of that observed for control chromosomal markers. The loss of resistance was enhanced by ethidium bromide treatment, but no chloramphenicol-resistant mutant was isolated by mutagenesis of a "cured" clone or naturally susceptible isolates. All attempts to isolate plasmid deoxyribonucleic acid as covalently closed circular molecules from strain BM 6001 have been unsuccessful, but epidemiological evidence and the fact that the genes specifying chloramphenicol acetyltransferase synthesis are usually located on plasmids suggest that this marker may be plasmid-borne in S. pneumoniae.

Acetylation↗

Molecular studies and possible relatedness between R plasmids from groups B and D streptococci.

Resistance plasmids isolated from Streptococcus agalactiae (group B) and S. faecalis (group D) have been compared in regard to resistance markers, molecular weight, and DNA-DNA homology. Three of them (pIP501, pIP612, and pIP613) have been found to confer identical (or very similar) resistance patterns (erythromycin, lincomycin, and streptogramin B, respectively) and to have similar molecular weights (19.8 x 10(6), 22.7 x 10(6), and 17.6 x 10(6), respectively) and a high level of DNA-DNA homology in hybridization experiments (90 to 100%). These results are compatible with the view that these plasmids may derive from one common ancestor, and/or that they can be transferred between unrelated Streptococcus strains belonging to the same or different groups.

Anti-Bacterial Agents↗

Formation of HfrH-type donor cells as a result of integrative suppression by R-F recombinant plasmids.

Three recombinant plasmids, resulting from recombination between an R plasmid of the FI incompatibility group and the F of HfrH, were introduced in a temperature-sensitive dnaA mutant to isolate Hfr-type-donors. All of the temperature-insensitive clones isolated from two of the three recombinant plasmids had the same origin and transfer pattern as the parental HfrH strain.

Chromosomes, Bacterial↗

[Microheterogeneity of R plasmid coded beta-lactamases in analytical isoelectric focusing (author's transl)].

Analytical isoelectric focusing of plasmid-coded beta-lactamases in polyacrylamide gels gives rise to several satellite bands in addition to one major active band (microheterogeneity). When the R plasmids are transferred by conjugation into 4 different bacterial species (Escherichia coli, Klebsiella pneumoniae, Salmonella typhi-murium, Proteus mirabilis) the position of the main band remains constant, but the number and position of the satellite bands are changed. Furthermore, the minimal inhibitory concentrations of several beta-lactamines are also changed when the plasmids are transferred between these different host strains. These facts suggest that the bacterial host might modify the physical structure and the activity of plasmid-determined beta-lactamases. They also suggest that identification of beta-lactamases and comparison of their isoelectric focusing patterns should always be carried out using the same bacterial host strain.

Amidohydrolases↗

New plasmid-mediated nucleotidylation of aminoglycoside antibiotics in Staphlococcus aureus.

A wild-type strain of Staphylococcus aureus, which inactivates a wide variety of aminoglycosides (except the gentamicin components), has been found to harbor a plasmid (RAp01) that mediates the biosynthesis of a nucleotidyltransferase. This enzyme modifies the 4'-hydroxy function of these antibiotics. The plasmid has been studied, the enzyme responsible for this resistance pattern has been isolated by affinity chromatography, and its kinetics and physicochemistry have been characterized. The target of this enzyme has also been located by demonstrating the structure of one inactivated compound, 4'-(O)-adenylyltobramycin.

Aminoglycosides↗

R plasmids in Streptococcus agalactiae (group B).

Two plasmids determining resistance to tetracycline (RIP500) and to chloramphenicol, erythromycin, lincomycin, and pristinamycin I (RIP501) were isolated from a strain of Streptococcus agalactiae. The frequency-of-resistance loss is very low for RIP500 (<3 x 10(4)) but higher for RIP501 (the efficiency was dependent upon the curing agents and incubation temperature and varied between 0.5 and 96%). Derivatives susceptible to all drugs were also obtained. RIP500 and RIP501 have similar molecular weights (17.9 x 10(6) and 20 x 10(6), respectively) and represent different percentages of total deoxyribonucleic acid (0.4 and 4%, respectively). The number of copies of RIP500 and RIP501 per cell is different, and these plasmids are likely replicated under different kinds of control (stringent and/or relaxed). No plasmid deoxyribonucleic acid was found in a derivative of strain B96 susceptible to all drugs.

Centrifugation, Density Gradient↗

Genetic and physical studies of recombinant plasmids formed between an R plasmid of compatibility group FI and sex factor F of HfrH.

Recombinant plasmids between an R plasmid of the FI group (R162/3) and the sex factor F or HfrH were produced after the conjugal transfer of this R plasmid into HfrH. Three types of recombinant plasmids were identified after the mating of HfrH (R162/3) with recA and rec+ recipients. One specimen of each type (pIP218, pIP222, pIP226) was studied in this report. All three recombinant plasmids carry the same genetic information for resistance to antibiotics (CSSuT) retained from R162/3. pIP218 retained all the other properties from F of HfrH: derepression for pilus synthesis, mobilization of the chromosome for the proximally transferred HfrH genes (thr, leu, proA), interference with T7 propagation, and ability to be cured by acridine orange. pIP222 retained from F of HfrH the derepression for pilus synthesis and the same polarity of chromosome transfer (thr, leu, proA), while pIP226 retained the interference with T7 propagation and acridine orange curing. Physical studies revealed that replication control and/or recovery of F and pIP218 as covalent circles of deoxyribonucleic acid are similar, and are different from R162/3. The new plasmids are more likely the result of a substitutive recombination event than a fusion. We propose genetic maps of these recombinant plasmids, showing the unequal participation of the parental plasmids in their formation.

Acridines↗

Physical studies of a plasmid mediating tetracycline resistance and hydrogen sulfide production in Escherichia coli.

Tetracycline resistance and hydrogen sulfide production have been previously found to be plasmid-mediated in a naturally-occurring strain of Escherichia coli; both functions are specified by a single conjugative plasmid called pIP231(Te-H2S); pIP231DNA was isolated as covalently closed molecules in dye-buoyant density gradients. The base ratio of the DNA was found to be 50% GC by density gradient analysis. Electron micrographs of plasmid molecules showed a contour length of 20 +/- 2 mum (40 +/- 4 X 10(6) daltons). Between one and two copies of pIP231 molecules per host chromosome were found in E. coli K12.

Base Sequence↗

[Tetracycline resistance in Haemophilus influenzae is plasmid-linked].

A strain of Haemophilus influenzae resistant to tetracycline (MIC 16-32 mug/ml) was "cured" of tetracycline resistance by ethidium bromide. DNA centrifugation in cesium chloride + ethidium bromide density gradients showed the existence of a band of non-chromosomal DNA molecules in the original resistant strain. This band was not found in lysates of a strain "cured" of tetracycline resistance by ethidium bromide. These facts support the hypothesis that in this strain of Haemohpilus influenzae, tetracycline resistance is plasmid mediated.

Centrifugation, Density Gradient↗

A transferable kanamycin resistance plasmid isolated from Haemophilus influenzae.

Kanamycin resistance was transferred by conjugation from a Haemophilus influenzae strain into Escherichia coli K12, and between E. coli K12 strains in subsequent transfers. Covalently closed molecules of DNA were isolated by sedimentation analysis of the DNA of the resistant E. coli K12 strains. These facts support the hypothesis that kanamycin resistance is mediated by a transferable plasmid in this strain of H. influenzae.

Conjugation, Genetic↗

Incompatibility groups and the classification of fi - resistance factors.

Incompatibility between R factors has been reported by several authors, and four incompatibility groups have already been described by Datta and Hedges among Rfi(-) factors. The stability of 12 plasmids in pairs was studied after 116 crosses, and five new groups were found, designated 5, 6, 7, 8, and 9. Each plasmid studied belongs to one single group. Incompatibility between plasmids in pairs is a clear-cut phenomenon, is easy to observe, and can provide a reliable method for recognizing and classifying resistance factors, and for tracing their spread among bacterial species.

Anti-Bacterial Agents↗