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JAC Advance Access originally published online on November 25, 2003
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Journal of Antimicrobial Chemotherapy (2004) 53, 58-64
© 2004 The British Society for Antimicrobial Chemotherapy

Molecular analysis of florfenicol-resistant Escherichia coli isolates from pigs

Maren Blickwede and Stefan Schwarz*

Institut für Tierzucht, Bundesforschungsanstalt für Landwirtschaft (FAL), Höltystrasse 10, 31535 Neustadt-Mariensee, Germany

Received 29 August 2003, returned 6 September 2003; revised 23 September 2003; accepted 3 October 2003

Objectives: The aim of this study was to analyse florfenicol-resistant Escherichia coli isolates from pigs for the genetic basis of florfenicol resistance, and to compare these data with those previously determined for E. coli isolates from cattle and poultry.

Methods: Fourteen porcine E. coli isolates were included in this study and subjected to serotyping, plasmid profiling and macrorestriction analysis. MICs of florfenicol were determined by broth microdilution. The presence of the gene floR was confirmed by hybridization and PCR analysis. Transformation experiments were conducted to isolate florfenicol resistance plasmids. The floR region of a florfenicol resistance plasmid was cloned and sequenced.

Results: All florfenicol-resistant E. coli isolates exhibited MICs of florfenicol >128 mg/L and carried the floR gene. A single isolate had a floR-carrying plasmid of ~35 kb, designated pMBSF1. Sequence analysis identified the floR gene flanked by truncated transposase genes. Moreover, a truncated copy of Tn5393 with complete streptomycin resistance genes strA and strB was found upstream of the floR gene of pMBSF1. Chromosomally resistant E. coli isolates, which shared the same BlnI macrorestriction pattern, differed in their floR hybridization patterns.

Conclusion: The plasmid pMBSF1 is the smallest floR-carrying plasmid reported to date. Its floR region differed from those previously found in E. coli isolates from cattle. Variations in the RFLPs of chromosomal EcoRI fragments carrying floR in isolates that had the same macrorestriction pattern might suggest variable chromosomal integration sites.

Keywords: florfenicol resistance, floR gene, transposon Tn5393, transposon Tn1721, recombination

* Corresponding author. Tel: +49-5034-871-241; Fax: +49-5034-871-246; E-mail: stefan.schwarz{at}fal.de


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