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T Horaud

Publications and source records attributed to T Horaud.

At least 19 recordsLinked to original sources

Diversity of structures carrying the high-level gentamicin resistance gene (aac6-aph2) in Enterococcus faecalis strains isolated in France.

Of 24 high-level gentamicin-resistant clinical isolates of Enterococcus faecalis, 20 carried gentamicin resistance (Gmr) plasmids. The plasmids ranged from 65.0 to 80.0 kb in size. Three of these plasmids were nonconjugative, and 17 transferred by conjugation to an E. faecalis recipient at low frequency (10(-5) to 10(-6) transconjugants per donor). The remaining four strains had a nonconjugative chromosomal Gmr determinant. On the basis of restriction enzyme and DNA-DNA hybridization profiles, Tn4001-like alpha elements were located on the chromosome and three types of Tn4001-truncated structures, I, II, and III, were found to be carried by the Gmr plasmids. Structure I lacked IS256 in the right-hand flanking extremity of Tn4001. Structure II was the same as structure I except that it also had a partial deletion of IS256 in the left-hand flanking extremity of Tn4001. Structure III lacked both the right- and left-hand flanking extremities of Tn4001. One of the wild-type strains carried the Gmr determinant both on the chromosome, as a Tn4001-like alpha element, and on a conjugative plasmid, as a Tn4001-truncated type I structure.

Chromosome Mapping↗

Molecular typing of Enterococcus faecalis strains resistant to high levels of gentamicin and isolated in Romania.

Sixteen Enterococcus faecalis strains resistant to high levels of gentamicin, 15 of which were isolated in the same year in a Romanian hospital, harboured conjugative gentamicin resistance (Gm(r)) plasmids ranging from 55 to 85 kilobases. On the basis of restriction enzyme and DNA-DNA hybridization profiles of these plasmids, as well as of chromosomal SmaI macrorestriction and Tn916 hybridization patterns, clonal relationship was established for seven strains whereas the other strains were considered to be independent. Nine and seven of the Gm(r) plasmids carried Tn4001-like and Tn4001-truncated structures, respectively; the latter structures were truncated in the right-hand flanking extremity of the element.

Chromosomes, Bacterial↗

New tetracycline resistance determinants coding for ribosomal protection in streptococci and nucleotide sequence of tet(T) isolated from Streptococcus pyogenes A498.

An approach based on PCR has been developed to identify new members of the tet gene family in streptococci resistant to tetracycline and minocycline. Degenerate primers, corresponding to portions of the conserved domains of the proteins Tet(M), Tet(O), TeTB(P), Tet(Q), and Tet(S), all specifying the tetracycline-minocycline resistance phenotype, were used to selectively amplify DNA fragments within the coding sequences. Nine streptococcal strains which do not carry the genes tet(M), tet(O), tetB(P), tet(Q), or tet(S) were investigated. Four of them gave no detectable PCR products. The five remaining strains each yielded a PCR product of 1.1 kbp. DNA hybridization experiments showed that these putative Tet determinants fell into four new hybridization classes, of which one, Tet T, was further analyzed. The gene tet(T) was isolated from Streptococcus pyogenes A498, and the nucleotide sequence that was necessary and sufficient for the expression of tetracycline resistance in Escherichia coli was determined. The deduced Tet(T) protein consists of 651 amino acids. The protein most closely related to Tet(T) was Tet(Q), which has 49% identical amino acid residues. A phylogenetic analysis revealed that Tet T represents a novel branching order among the Tet determinants so far described.

Amino Acid Sequence↗

Chromosomal gentamicin resistance transposon Tn3706 in Streptococcus agalactiae B128.

Streptococcus agalactiae B128 is the only highly gentamicin-resistant group B streptococcal (GBS) strain described so far. This strain carries a chromosomal gentamicin resistance transposon, designated Tn3706, which is similar, if not identical, to the Tn4001 and Tn5281 transpons detected in Staphylococcus aureus and Enterococcus faecalis, respectively. Transposition of Tn3706 occurred onto the GBS plasmid pIP501 in two different loci of its 7.5-kb AvaII fragment carrying the genes for chloramphenicol and erythromycin resistance. Molecular analysis of pIP501 derivatives showed that Tn3706 is composed of a central fragment containing the aac6'-aph2" gene; this fragment is flanked by two tandemly repeated copies of IS256 at the 5' extremity of the resistance gene and a single inverted copy of IS256 at its 3' extremity. The two tandemly repeated copies of IS256 were separated by a 6-bp segment identical to that found, in the same orientation, in the IS256-aac6'-aph2" junction. The hybrid replicons pIP501::Tn3706 were found to be structurally unstable following conjugative transfer between GBS strains. Numerous individual copies of IS256 were detected in B128, but this insertion sequence was not found in the 11 wild-type, gentamicin-susceptible GBS strains studied.

Anti-Bacterial Agents↗

Plasmid-borne high-level resistance to gentamicin in Enterococcus hirae, Enterococcus avium, and Enterococcus raffinosus.

Enterococcus hirae, E. avium, and E. raffinosus isolated in Romania, Tunisia, and Portugal harbored plasmids pICC8, pIP1700, and pIP1701, respectively, encoding resistance to high levels of gentamicin (Gmr). The Gmr marker was carried on pIP1700 by a Tn4001-like element and on pICC8 and pIP1701 by Tn4001-truncated structures. pICC8 carried, in addition to Gmr, chloramphenicol, erythromycin, and tetracycline-minocycline (TetM) resistance determinants. The gene tetM of pICC8 was carried on a Tn916-like element.

Drug Resistance, Microbial↗

Genetic and molecular studies of a composite chromosomal element (Tn3705) containing a Tn916-modified structure (Tn3704) in Streptococcus anginosus F22.

The plasmid-free Streptococcus anginosus F22 contained a conjugative element, Tn3705, encoding resistance to erythromycin (Emr) and tetracycline-minocycline (Tcr-Mnr). We mapped a chromosomal region (> 52 kb) of F22, corresponding to the internal part of Tn3705. Molecular analysis of Tn3705 revealed it to be a composite structure: it included in its central part a transposon designated Tn3704 (20.3 kb +/- 0.5 kb), which had a modified structure in comparison with that of Tn916 and on which the Emr Tcr-Mn4 markers were localized. Tn3705 inserted from F22 into the chromosome of various streptococcal transconjugants as well as that of Enterococcus faecalis transconjugants without changing its structure. In contrast, from the chromosome of an E. faecalis::Tn3705 transconjugant only Tn3704 inserted, at various sites, into another E. faecalis chromosome. Sugar fermentations occurred after the insertion of Tn3704 into the chromosome of an asaccharolytic E. faecalis strain. Transposition of only Tn3704 from the chromosome of E. faecalis::Tn3705 onto pIP964, an E. faecalis hemolysin plasmid, yielded two different pIP964 derivatives. The size of the entire element Tn3705 was estimated to be about 70.0 kb by pulsed-field electrophoresis.

Chromosomes, Bacterial↗

Diversity of chromosomal genetic elements and gene identification in antibiotic-resistant strains of Streptococcus pneumoniae and Streptococcus bovis.

Antibiotic-resistant Streptococcus pneumoniae (26 strains) and Streptococcus bovis (28 strains), devoid of R plasmids, were examined for DNA-DNA homology to Tn916 and Tn3701. Tn916-like structures were found in 17 S. pneumoniae and 21 S. bovis strains. Tn916-modified structures were present in 6 S. pneumoniae and 2 S. bovis strains. Two strains of each species carried elements having a Tn3701-like composite structure. All these elements were chromosome-borne. No chromosomal elements were detected in 1 S. pneumoniae and 3 S. bovis strains.

Chromosomes, Bacterial↗

Study of heterogeneity of chloramphenicol acetyltransferase (CAT) genes in streptococci and enterococci by polymerase chain reaction: characterization of a new CAT determinant.

An assay based on the utilization of degenerate primers that enable enzymatic amplification of an internal fragment of cat genes known to be present in gram-positive cocci was developed to identify the genes encoding chloramphenicol resistance in streptococci and enterococci. The functionality of this system was illustrated by the detection of cat genes belonging to four different hydridization classes represented by the staphylococcal genes catpC221, catpC194, catpSCS7, and the clostridial gene catP, and by the characterization of a new streptococcal cat gene designated catS. A sequence related to the clostridial catQ gene, which was present in one streptococcal strain, was not detected by this assay. These results reveal that these six cat genes account for chromosomal-borne chloramphenicol resistance in 12 group A, B, and G streptococci tested. By contrast, only three of these six cat genes (catpC221, catpC194, and catpSCS7) were detected on the 10 enterococcal plasmids studied here that encode resistance to chloramphenicol.

Amino Acid Sequence↗

Identification of new sex pheromone plasmids in Enterococcus faecalis.

We describe the identification of the following new sex pheromone plasmids in Enterococcus faecalis: a haemolysin-bacteriocin plasmid, pIP964; three R plasmids, pIP1017, pIP1438 and pIP1440; and two cryptic conjugative plasmids, pIP1141 and pMV120. The identification was based on the formation of cell aggregates on filter membranes during conjugation, on efficient transfer in broth matings, and on a positive clumping reaction of cells carrying these plasmids. In addition these plasmids hybridized with DNA probes specific for sex pheromone-induced structural genes encoding surface proteins required for conjugative transfer of the plasmids.

DNA Probes↗

Nucleotide sequence of the chloramphenicol resistance determinant of the streptococcal plasmid pIP501.

We have sequenced the chloramphenicol resistance determinant (cat) of plasmid pIP501 from Streptococcus agalactiae to investigate its relationship with other cognate cat determinants. Sequence analysis revealed that it exhibits a high degree of similarity with the cat genes of plasmids pC221 and pUB112 from Staphylococcus aureus and pSCS1 from Staphylococcus intermedius. These genes, however, display several differences in their regulatory and coding regions. These results demonstrate that the cat determinant of plasmid pIP501 belongs to the pC221 subgroup of CAT variants.

Amino Acid Sequence↗