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Edwardsiella piscicida  is a pathogenic bacterium responsible for significant losses in important wild and cultured fish species.  E. piscicida  strain MS-18-199 recovered from a diseased hybrid catfish from East Mississippi and showed resistance to florfenicol, chloramphenicol, oxytetracycline, doxycycline, erythromycin, tetracycline, azitromycin, spectinomycin, sulfonamide, and bacitracin. To explore the mechanisms of resistance in  E. piscicida  strain MS-18-199, genomic DNA was extracted and subjected to whole genome sequencing (WGS) using a combination of long (Oxford Nanopore) and short (Illumina) reads. The genome of strain MS-18-199 revealed a novel plasmid named pEPMS-18199. The 117,448 bp plasmid contains several antimicrobial resistance (AMR) elements/genes, including florfenicol efflux pump ( floR ), tetracycline efflux pump ( tetA ), tetracycline repressor protein ( tetR ), sulfonamide resistance ( sul2 ), aminoglycoside O-phosphotransferase  aph(6) -Id ( strB ), and aminoglycoside O-phosphotransferase  aph(3) -Ib ( strA ). Two genes,  arsA  and  arsD , that encode protein components related to transport/resistance to arsenic were also found in pEPMS-18199. In addition, pEPMS-18199 carried twelve conjugative transfer genes ( tra ), eight transposases and insertion elements, two plasmid stability proteins, two replication proteins, and three partitioning proteins ( par  system). Results from mobilization and stability experiments revealed that pEPMS-18199 is highly stable in the host cell and could be transferred to  Escherichia coli  and  Edwardsiella ictaluri  by conjugation. To our knowledge, this is the first detection of a multidrug resistance (MDR) conjugative plasmid in  E. piscicida  in the United States. Careful tracking of this plasmid in the aquaculture system is warranted. Knowledge regarding the molecular mechanisms of AMR in aquaculture is important for antimicrobial stewardship.

Characterization of a Novel Conjugative Plasmid in Edwardsiella piscicida Strain MS-18-199