Molecular Epidemiology of Antibiotic Resistance Genes in Multiresistant EpidemicSalmonella typhimuriumDT 104

Abstract

The epidemiology of antibiotic resistance genes in epidemic multiresistant S. typhimurium DT 104 of human and animal origin was investigated. DNA prepared from 45 human and 21 animal strains isolated between 1984 and 1997, including eight isolated in other European countries, the USA, Trinidad, and South Africa and resistant to ampicillin, chloramphenicol, streptomycin, sulphonamides, spectinomycin, tetracyclines (R-type ACSSuSpT) were examined for the presence of integrons by PCR. Integron hot spots were observed in all strains conferring resistance to ACSSuSpT in two copies, determined by two discrete bands of approximately 1.0 and 1.2 kb. Direct nucleotide sequencing of the individual amplicons of selected strains indicated that the 1.0 kb gene product was ant (3′′)-Ia, responsible for resistance to streptomycin and spectinomycin; the 1.2 kb amplicon contained the gene bla_PSE-1, encoding the β-lactamase PSE-1 (CARB-2). Both integrons were encoded on a single XbaI macrorestriction fragment of approximately 10 kb. All isolates of DT 104 of this resistance phenotype contained the same inserted gene cassettes, irrespective of source and country of origin, supporting the suggestion of the spread of an epidemic clone. Sequence analysis of the quinolone resistance determining region (QRDR) of gyrA of 15 multiresistant strains conferring additional resistance to nalidixic acid and ciprofloxacin (R-type ACSSuSpTNxCp) identified two discrete base substitutions at codon Asp-87. Conversion of Asp-87 → Asn was most commonly observed, in 7/10 human and 4/5 animal isolates, suggesting that this codon plays a major role in the development of ciprofloxacin resistance in multiresistant S. typhimurium DT 104.