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C Werckenthin

Publications and source records attributed to C Werckenthin.

6 recordsLinked to original sources

Molecular analysis of naturally occuring ermC-encoding plasmids in staphylococci isolated from animals with and without previous contact with macrolide/lincosamide antibiotics.

A total of 16 epidemiologically unrelated macrolide-resistant staphylococcal isolates of various animal origins were investigated for the molecular basis of macrolide resistance with respect to previous contact of their host animals with macrolides and lincosamides. All isolates carried ermC-encoding plasmids of 2.3-4.0 kbp. The eight plasmids of staphylococci from animals which had not received macrolides or lincosamides showed inducible ermC gene expression and did not exhibit alterations in the ermC regulatory region. The remaining eight plasmids expressed the ermC gene constitutively. Six of these plasmids were from staphylococci from animals which had received tylosin or spiramycin as feed additives or lincomycin for therapeutic purposes. All constitutively expressed ermC genes revealed either sequence deletions or sequence duplications in their ermC regulatory region, as detected by a PCR assay and by sequence analysis. These sequence deletions and duplications found in naturally occurring plasmids corresponded closely to the mutations seen in the ermC-encoding plasmids after growth of an inducibly resistant strain in the presence of non-inducing macrolides or lincosamides under in vitro conditions.

Amino Acid Sequence

A novel plasmid from Staphylococcus epidermidis specifying resistance to kanamycin, neomycin and tetracycline.

The naturally occurring plasmid pSTS7 from Staphylococcus epidermidis mediated resistance to tetracycline via a tetL gene and to kanamycin and neomycin via an aadD gene. Plasmid pSTS7 showed partial restriction map and sequence homology to the previously described tetracycline resistance plasmid pNS1981 from Bacillus subtilis and to the kanamycin/neomycin/bleomycin resistance plasmid pUB110 from S. aureus. Sequence analysis of the regions flanking the two resistance genes in pSTS7 led to the identification of a novel site for interplasmid recombination which could explain the derivation of pSTS7 from the incompatible pNS1981- and pUB110-like parental plasmids under tetracycline-selective pressure.

Amino Acid Sequence

Integration of pT181-like tetracycline resistance plasmids into large staphylococcal plasmids involves IS257.

Four large staphylococcal plasmids ranging in size from 31 to 82 kbp have been shown to mediate tetracycline resistance via an integrated copy of the tet(K)-encoding plasmid pT181 which was flanked by copies of the insertion element IS257. In two cases, IS257 elements interrupted the repC reading frame of pT181 and an 8-bp sequence from within the repC gene was duplicated at the interrupted site. In the third plasmid, the IS257 elements interrupted the pT181 DNA immediately upstream of the repC coding sequence with an 8-bp duplication. In the fourth case, the IS257 elements flanked a pT181-like plasmid with one IS257 in the repC coding sequence and the other within the recombinase (pre) coding sequence, so that a section of the pT181 sequence was deleted. All four integration sites detected in this study differ from those previously described for the IS257-mediated integration of pT181-like plasmids into large plasmids or into the chromosomal DNA.

Blotting, Southern

Macrolide-lincosamide-streptogramin B resistance in Staphylococcus lentus results from the integration of part of a transposon into a small plasmid.

The 8.0-kb macrolide-lincosamide-streptogramin B resistance plasmid pSES20 from Staphylococcus lentus harbored part of a Tn917-like transposon including the left terminal repeat, a gene almost identical to ermB, and its regulatory region, as well as the internal direct repeat. Homology between pSES20 and Tn917 ended at a sequence closely related to those of the resolution sites of Tn917 and Tn552 and staphylococcal recombination sites.

Amino Acid Sequence

Chloramphenicol resistance in Staphylococcus intermedius from a single veterinary centre: evidence for plasmid and chromosomal location of the resistance genes.

A total of seven Staphylococcus intermedius cultures isolated from cases of canine pyoderma were investigated for the genetic basis of chloramphenicol resistance (Cmr). All of these S. intermedius isolates mediated Cmr via the expression of the Cm-inactivating enzyme chloramphenicol acetyltransferase (CAT); the respective cat genes were found to be located on small multicopy plasmids of 3.1 to 4.1 kb in four of the seven cultures. The four Cmr plasmids, designated pSCS20-23, differed upon restriction endonuclease mapping. Hybridization experiments identified all of them to belong to the pC221-family of staphylococcal Cmr plasmids. The expression of all four plasmid-encoded cat genes was inducible with chloramphenicol. The remaining three S. intermedius isolates also harboured an inducible cat gene of the pC221-type which, however, was found to be located in the chromosomal DNA. These differences in the subcellular localisation and consequently in the number of cat gene copies per S. intermedius cell had no influence on the MIC values of Cm exhibited by the respective S. intermedius isolates.

Animals

Insertion elements in Staphylococcus intermedius.

Staphylococcus intermedius cultures from dogs, pigeons, horses and mink were investigated for the prevalence of the insertion elements IS256 and IS257 in relation to their antibiotic resistance. Copies of IS256 could not be detected in any of the Staph. intermedius isolates tested whereas single copies of IS257 occurred in the isolates from dogs and horses. The mink strains did not harbour IS257 elements, whereas Staph. intermedius isolates from pigeons carried multiple copies of IS257 as predicted from the hybridization patterns obtained with a gene probe derived from the internal part of the IS257-encoded transposase gene. Independently of the origin of the Staph. intermedius isolates, all IS257 copies were found to be located in the chromosomal DNA. The large number of chromosomal IS257 copies in the pigeon strains might help to explain chromosomal multiresistance in many of those strains.

Animals