title: Distribution and in-vitro transfer of tetracycline resistance determinants in clinical and aquatic Acinetobacter strains.
abstract: Following characterisation by phenotypic tests and amplified ribosomal DNA restriction analysis (ARDRA), 50 tetracycline-resistant (MIC > or = 16 mg/L) Acinetobacter strains from clinical (n = 35) and aquatic (n = 15) samples were analysed by PCR for tetracycline resistance (Tet) determinants of classes A-E. All the clinical strains were A. baumannii; most (33 of 35) had Tet A (n = 16) or B (n = 17) determinants, and only two did not yield amplicons with primers for any of the five tetracycline resistance determinants. The aquatic strains belonged to genomic species other than A. baumannii, and most (12 of 15) did not contain determinants Tet A-E. Strains negative for Tet A-E were also negative for Tet G and M; further analysis of two aquatic strains with specific primers for Tet O and Tet Y and degenerate primers for Tet M-S-O-P(B)-Q also showed negative results. Transfer of tetracycline resistance was tested for 20 strains with three aquatic Acinetobacter strains and Escherichia coli K-12 as recipients. Transfer of resistance was demonstrated between aquatic strains from distinct ecological niches, but not from clinical to aquatic strains, nor from any Acinetobacter strain to E. coli K-12. Most transconjugants acquired multiple relatively small plasmids (<36 kb). Transfer did not occur when DNA from the donor strains was added to the recipient cultures and was not affected by deoxyribonuclease I, suggesting a conjugative mechanism. It is concluded that Tet A and B are widespread among tetracycline-resistant A. baumannii strains of clinical origin, but unknown genetic determinants are responsible for most tetracycline resistance among aquatic Acinetobacter spp. These differences, together with the inability of clinical strains to transfer tetracycline resistance in vitro to aquatic strains, contra-indicate any important flow of tetracycline resistance genes between clinical and aquatic acinetobacter populations.
authors.forename: L
authors.initials: L
authors.lastname: Guardabassi
authors.affiliation: Department of Veterinary Microbiology, The Royal Veterinary and Agricultural University, 1870 Frederiksberg C., Denmark; *Department of Infectious Diseases, Leiden University Medical Centre, 2300 RC Leiden, Netherlands; and †Section of Biosafety and Biotechnology, Scientific Institute Public Health-Louis Pasteur, B-1050 Brussels, Belgium.
authors.forename: L
authors.initials: L
authors.lastname: Dijkshoorn
authors.affiliation: Department of Veterinary Microbiology, The Royal Veterinary and Agricultural University, 1870 Frederiksberg C., Denmark; *Department of Infectious Diseases, Leiden University Medical Centre, 2300 RC Leiden, Netherlands; and †Section of Biosafety and Biotechnology, Scientific Institute Public Health-Louis Pasteur, B-1050 Brussels, Belgium.
authors.forename: J-M
authors.initials: JM
authors.lastname: Collard
authors.forename: J E
authors.initials: JE
authors.lastname: Olsen
authors.forename: A
authors.initials: A
authors.lastname: Dalsgaard
mesh_terms: D000150:Acinetobacter / Q000145:classification / Q000187:drug effects* / Q000235:genetics; D000151:Acinetobacter Infections / Q000188:drug therapy* / Q000382:microbiology; D000900:Anti-Bacterial Agents / Q000494:pharmacology* / Q000627:therapeutic use; D017931:DNA Primers / Q000737:chemistry; D004262:DNA Restriction Enzymes / Q000737:chemistry; D004275:DNA, Ribosomal / Q000737:chemistry; D004352:Drug Resistance, Microbial; D004587:Electrophoresis, Agar Gel; D018014:Gene Transfer Techniques; D006801:Humans; D010957:Plasmids / Q000737:chemistry; D016133:Polymerase Chain Reaction; D013752:Tetracycline / Q000494:pharmacology* / Q000627:therapeutic use; D013753:Tetracycline Resistance / Q000235:genetics*; D014871:Water Microbiology
chemical_list: D000900:Anti-Bacterial Agents; D017931:DNA Primers; D004275:DNA, Ribosomal; D004262:DNA Restriction Enzymes; D013752:Tetracycline