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Mechanisms of Resistance

Class 1 Integron-Borne Multiple-Antibiotic Resistance Carried by IncFI and IncL/M Plasmids in Salmonella enterica Serotype Typhimurium

Fabio Tosini, Paolo Visca, Ida Luzzi, Anna Maria Dionisi, Cristina Pezzella, Andrea Petrucca, Alessandra Carattoli
Fabio Tosini
Laboratory of Cellular Biology 1 and
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Paolo Visca
Laboratory of Bacteriology and Medical Mycology, 2
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Ida Luzzi
Laboratory of Bacteriology and Medical Mycology, 2
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Anna Maria Dionisi
Laboratory of Bacteriology and Medical Mycology, 2
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Cristina Pezzella
Laboratory of Bacteriology and Medical Mycology, 2
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Andrea Petrucca
Istituto Superiore di Sanità Rome, and Institute of Microbiology, University of Rome, “La Sapienza,” 3 Rome, Italy
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Alessandra Carattoli
Laboratory of Bacteriology and Medical Mycology, 2
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DOI: 10.1128/AAC.42.12.3053
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    Fig. 1.

    Plasmids in S. enterica serotype Typhimurium strains. (A) Plasmid DNA in S. enterica serotype Wien WZM6 (lane 1) and in six prototypic isolates of S. entericaserotype Typhimurium, strains 44 (lane 2), 516 (lane 3), 252 (lane 4), 298 (lane 5), 202 (lane 6), and 341 (lane 7). Plasmids were separated by agarose gel electrophoresis and stained with ethidium bromide. Replicas of the electropherograms shown in panel A were blotted onto nylon membrane filters and hybridized with the following probes:rep FIA (B), rep FIB (C),rep L/M (D), and an integrase gene (intI1) (E). Plasmid pACYC184::Tn21(lane 8) was used as positive control for the presence of theintI1 gene.

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    Fig. 2.

    Integrons found on plasmids carried by S. enterica serotype Typhimurium. (A) Plasmids were separated by agarose gel electrophoresis and were visualized by ethidium bromide staining. Lane 1, S. enterica serotype Typhimurium 202 donor strain; lane 2, Cm-type E. coli exconjugant carrying the IncFI replicon; lane 3, Ak-type E. coli exconjugant carrying the IncL/M replicon. (B) PCR analysis of integron-borne gene cassettes amplified with the 5′CS and 3′CS primers (Table 1). DNA templates were from the S. enterica serotype Typhimurium 202 (lane 1) and from Ak-type and Cm-type exconjugants (lanes 2 and 3, respectively). In-t1, In-t2, and In-t3 integrons are indicated as 1, 2, and 3, respectively, on the left of the figure. Molecular size standards (lane 4; in kilobases) are indicated on the right.

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    Fig. 3.

    Structural organization of gene cassettes in S. enterica serotype Typhimurium integrons. Integron-borne gene cassettes are represented as open boxes; black boxes are the 59-base elements (not in scale). The positions of an integrase gene (intI1), Pant promoter, a sulfonamides resistance gene (sul-1), and the 5′CS and 3′CS primers are indicated by arrows. Restriction sites for BamHI (B), PvuII (P), and Rsa I (R) are indicated.

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  • Table 1.

    Primers used for integron detection by PCR

    PrimerDNA sequencePosition in submitted sequenceAccession no.
    5′CS5′-GGCATCCAAGCAGCAAG-3′1190–1206 M73819
    3′CS5′-AAGCAGACTTGACCTGA-3′1342–1326 M73819
    intI2A5′-ATGTCTAACAGTCCATTTTTAAATTCTA-3′1914–1886 AJ002782
    intI2B5′-AAATCTTTAACCCGCAAACGC-3′1475–1495 AJ002782
    intI3A5′-GTGGCGCAGGGTGTGGAC-3′194–211 D50438
    intI3B5′-ACAGACCGAGAAGGCTTATG-3′959–938 D50438
  • Table 2.

    Antibiotic resistance phenotypes of S. enterica serotype Typhimurium donor strains and E. coli exconjugants, transferred plasmids, and type of integrons found on plasmids

    Salmonella R typeNo. of strainsDonor strainExconjugant R typeFrequency of transferTransferred plasmidIntegron(s)
    ApCmKmSmSuTpTeNa5366ApCmKmSmSuTpTe5 × 10−5 140-kb IncFIIn-t1,In-t2
    ApCmKmSmSuTpNa9341ApCmKmSmSuTp7 × 10−5 140-kb IncFIIn-t1,In-t2
    ApCmKmSmSuTpTe15252ApCmKmSmSuTe6 × 10−5 140-kb IncFIIn-t1,In-t2
    ApCmKmSmSuTpTeGmAk8202ApKmSuTeGmAk1 ×10−6 100-kb IncL/MIn-t3
    202ApCmKmSmSuTpTe7 × 10−5 140-kb IncFIIn-t1,In-t2
    298ApKmSuGmAk7 × 10−7 100-kb IncL/MIn-t3
    298ApCmKmSmSuTpTe5 × 10−5 140-kb IncFIIn-t1,In-t2
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Class 1 Integron-Borne Multiple-Antibiotic Resistance Carried by IncFI and IncL/M Plasmids in Salmonella enterica Serotype Typhimurium
Fabio Tosini, Paolo Visca, Ida Luzzi, Anna Maria Dionisi, Cristina Pezzella, Andrea Petrucca, Alessandra Carattoli
Antimicrobial Agents and Chemotherapy Dec 1998, 42 (12) 3053-3058; DOI: 10.1128/AAC.42.12.3053

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Class 1 Integron-Borne Multiple-Antibiotic Resistance Carried by IncFI and IncL/M Plasmids in Salmonella enterica Serotype Typhimurium
Fabio Tosini, Paolo Visca, Ida Luzzi, Anna Maria Dionisi, Cristina Pezzella, Andrea Petrucca, Alessandra Carattoli
Antimicrobial Agents and Chemotherapy Dec 1998, 42 (12) 3053-3058; DOI: 10.1128/AAC.42.12.3053
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