cThe nucleotide sequences of three IncU plasmids from Aeromonas spp. isolated from ornamental fish are described. They had a typical IncU backbone for plasmid replication and maintenance functions, but conjugative transfer modules were disrupted. The gene qnrS2 was inserted into mpR as a mobile insertion cassette. Novel Tn3 family transposons carrying putative toxin-antitoxin and plasmid stability genes were identified. The study demonstrates high plasticity of IncU plasmids from aquatic environments.
Aeromonas is a ubiquitous microorganism distributed in water ecosystems and includes species causing various kinds of infections in fish and humans. Antibiotic resistance in Aeromonas in aquatic environments is associated especially with plasmids of the IncU family. Most IncU plasmids described in this genus to date share a highly conserved backbone represented by genes for plasmid replication, stability, and conjugative transfer and one variable region for resistance genes to antibiotics, such as tetracyclines, sulfonamides, trimethoprim, streptomycin, and chloramphenicol (1-6). Quinolones are broad-spectrum antimicrobial agents widely used in human and veterinary medicine, and they are among the most widely used antibiotics in aquacultures (7). Aeromonas isolates harboring quinolone resistance (PMQR) genes on IncU plasmids have been recently reported in river water, lake water, and fish (2,3,6,8), highlighting the role of this plasmid family in dissemination of clinically relevant resistance mechanisms in a wide range of aquatic environments.In our previous study, IncU plasmids carrying qnrS2 and aac(6=)-Ib-cr genes in Aeromonas spp. from ornamental fish originating from various geographic locations were identified (8). The aim of this study was to elucidate the structure and features of three representative IncU plasmids carrying PMQR genes by sequencing and bioinformatic analysis. At present, complete nucleotide sequences of nine IncU plasmids are available in the GenBank database. Four of them, plasmids pRA3 (GenBank accession no. DQ401103), pFBAOT6 (GenBank accession no. CR376602), pAC3 (GenBank accession no. KM204147), and pP2G1 (GenBank accession no. HE616910), originate from Aeromonas spp., and only the last two plasmids carry PMQR genes (updated on 22 July 2015).The three sequenced IncU plasmids pAH6, pAH227, and pASCH21 were nonconjugative and varied in size (20 to 40 kb). Plasmid pAH6 (ϳ20 kb) originated from Aeromonas hydrophila from a koi carp bred in the Czech Republic. It has been identified as a highly diffused plasmid in isolates of A. hydrophila and Aeromonas sobria from Czech koi carps and tropical fish imported from Asia (8). The other two plasmids, pAH227 (ϳ40 kb) and pASCH21 (ϳ35 kb), carrying qnrS2, originated from A. hydrophila from a koi carp from the Czech Republic and A. sobria from a tropical fish from Thailand, respectively. Plasmids were transferred to chemically competent Escherichia coli DH5␣ cells by transformation followed by selection of the transformants on Luria-Bertani agar wi...