Complete Sequence of the FII Plasmid p42-2, Carrying bla _CTX-M-55 , oqxAB , fosA3 , and floR from Escherichia coli

Abstract

dWe sequenced a novel conjugative multidrug resistance IncF plasmid, p42-2, isolated from Escherichia coli strain 42-2, previously identified in China. p42-2 is 106,886 bp long, composed of a typical IncFII-type backbone (ϳ54 kb) and one distinct acquired DNA region spanning ϳ53 kb, harboring 12 antibiotic resistance genes [bla CTX-M-55 , oqxA, oqxB, fosA3, floR, tetA(A), tetA(R), strA, strB, sul2, aph(3=)-II, and ⌬bla TEM-1 ]. The spread of these multidrug resistance determinants on the same plasmid is of great concern and, because of coresistance to antibiotics from different classes, is therapeutically challenging. Widespread antibiotic resistance poses an enormous threat for human and animal health worldwide (1). This problem has been exacerbated in recent years with the emergence of multidrug resistance (MDR) plasmids conferring resistance to most classes of antimicrobials (2). Plasmids of the incompatibility F (IncF) group, representing one of the most frequently encountered plasmid types (3), have frequently been associated with MDR phenotypes, including extended-spectrum ␤-lactamases (ESBLs) and plasmid-mediated quinolone resistance (PMQR) genes (4, 5). In a previous study of ours, IncF plasmids recovered from Escherichia coli strains of food-producing and companion animals were investigated, and most of them carried numerous resistance determinants, such as bla CTX-M , rmtB, oqxAB, and floR (4, 6). The spread of these multiresistance plasmids has prompted worldwide concern because of coresistance to multiple antimicrobial agents that facilitates the survival of bacteria under the selective pressure of antibiotics. Herein, we analyzed a common subtype IncF plasmid, F33:AϪ:BϪ (FII, FIA, FIB [FAB] formula); this plasmid, designated p42-2, contains 12 different resistance genes and was fully sequenced, and the data were compared with those of other IncF plasmids.E. coli 42-2 was recovered from the feces of a healthy duck in Guangzhou, China. Conjugation was performed by mixing E. coli 42-2 and E. coli C600 in a liquid medium and isolating for E. coli C600 (p42-2) by selection on MacConkey agar containing streptomycin (1,000 mg/liter) and cefotaxime (2 mg/liter) as previously described (4, 6). p42-2 was extracted from the E. coli C600 transconjugant using a commercial kit (Qiagen midikit; Qiagen, Germany). Sequencing of p42-2 was performed on an Illumina IIx genome analyzer with a 500-bp paired-end library (approximately 100 million available reads, 935-fold genome coverage) and a 2,000-bp paired-end library (ϳ337 million available reads, 3,150-fold genome coverage). These raw data were assembled by SOAPdenovo (7). Gene prediction and annotation were performed using RAST tools (8). The sequence comparison and map generation were performed using BLAST (http://blast.ncbi.nlm.nih.gov) and Easyfig version 2.1 (9).Plasmid p42-2 confers resistance to ampicillin, chloramphenicol, kanamycin, streptomycin, sulfonamides/trimethoprim, tetracycline, olaquindox, fosfomycin, cephalosporin, and florfenicol. Because of its ...

Complete Sequence of a F33:A-:B- Conjugative Plasmid Carrying the oqxAB, fosA3, and blaCTX-M-55 Elements from a Foodborne Escherichia coli Strain

“…In such fragment, the orf2 and orf3 downstream of fosA3 are orientated in opposite directions, and that the orf1 was truncated due to insertion of an additional copy of IS 26 ; such genetic arrangement differs from that commonly observed in other plasmids ( Figure 2B ). Compared to another IncFII plasmid, p42-2, which harbored the same batch of resistance elements as that in pE80, sequence analysis revealed that the genetic arrangement varied significantly between two plasmids (Yang et al, 2016). The ∼26.5 kb IS 26 -flanked sequence comprising the oqxAB locus and some other accessory genes in pE80 was found to be oppositely oriented in p42-2.…”

“…Except for the original oqxAB -bearing plasmid, pOLA52, which was first recovered in E. coli , information regarding the mobile elements that mediate the transmission of oqxAB among strains of the Enterobacteriaceae family remains scarce. Recently, the complete sequence of an IncFII plasmid carrying oqxAB , fosA3 , bla CTX-M-55, and floR from an E. coli of veterinary origin has been described (Yang et al, 2016). In this study, we reported the complete sequence of a F33:A-:B- conjugative plasmid carrying the oqxAB , fosA3 , floR , and bla CTX-M-55 genes harbored by an E. coli strain isolated from chicken meat from a retail store in Shenzhen, China.…”

The nature and epidemiology of OqxAB, a multidrug efflux pump

“…Many reports emphasized the co-spread of oqxAB with other resistance genes, virulence genes and heavy metal resistance genes in E.coli . The sequences of several plasmids from foodborne E. coli , such as pEC012 [51], pE80 [52] and p42–2 [53] had been published in the Genebank. These conjugative plasmids harbored multiple resistance determinants, including oqxAB , bla CTX-M-65 , rmtB , fosA3 and floR , indicating a therapeutic challenge due to co-selection by various antibiotics and thus compromise the effectiveness of current antimicrobial strategies.…”

Evolution and Comparative Genomics of F33:A−:B− Plasmids Carrying bla CTX-M-55 or bla CTX-M-65 in Escherichia coli and Klebsiella pneumoniae Isolated from Animals, Food Products, and Humans in China

“…In addition, F33:A−:B− or IncN-F33:A−:B− plasmids were responsible for the dissemination of fosA3 and bla CTX-M-55/-65 genes in E. coli from chickens in China and pigs, chickens, and dairy cows in northeast China ( 9 – 11 ). Furthermore, oqxAB colocated with bla CTX-M-55 on F33:A−:B− and IncN-F33:A−:B− plasmids was identified in food-producing animals, chicken meat, and humans in China ( 12 – 15 ). Recently, F33:A−:B− plasmids were also described as carriers of bla NDM-1 from porcine E. coli isolates in China ( 16 ).…”