Conceived and designed the experiments: JR TW. Performed the experiments: JR EC TW WF PM DW MR. Analyzed the data: JR MR DR TW WF ME PM DW. Contributed reagents/materials/analysis tools: JR EC LR DW TW PM DW MM JL TC MR LL JH. Wrote the paper: JR TW WF. Other: Edited Manuscript: EC TC DW PM.
The authors have declared that no competing interests exist.
Antimicrobial resistance in
Recently there also has been a rapid worldwide emergence of MDR foodborne bacterial pathogens
Here we report the complete sequence and comparative analysis of three nearly identical large (>150 kb) MDR plasmids isolated from
Comparative analysis of the complete DNA sequence of the
Nucleotide composition is represented on each of the outer circles and PCR primers used in this study are indicated in the inner circle. Antimicrobial determinants are colored red and labeled with gene names as follows: sulfonamides (
Backbone | ||||
Resistance phenotypes | Su | ACKSSpTMSul | ACSSulTCfAxG | STTmSul |
β-lactams | - | - | ||
Aminoglycosides/Aminocyclitols | - | |||
Tetracyclines | - | |||
Phenicols | - | - | ||
Quaternary ammonium compounds | - | - | ||
Sulfonamides | ||||
Trimethoprim | - | - | - |
Abbreviations used for antibiotics: A, ampicillin; C, chloramphenicol; S, streptomycin; Sul, sulfonamide; T, tetracycline; Cf, cephalothin, Ax, ceftriaxone; G, gentamicin; Sp, spectinomycin; M, minocycline; Tm, trimethoprim.
While MDR
Strain | Organism | State | Year | Source | Antimicrobial resistance phenotype | Transfer | |||||||||||||||||||||||||
A | C | S | Su | T | Ti | G | K | Ag | Ax | N | Ak | Cp | Sxt | ||||||||||||||||||
30034 | CA | 2003 | Turkey | A | C | S | Su | T | Ti | - | - | Ag | - | N | - | - | - | 1 | 2 | 3 | 4 | 5 | - | - | - | - | 10 | 11 | 12 | - | |
32412 | CT | 2003 | Chicken | A | - | - | Su | T | Ti | - | - | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | - | 11 | 12 | 8.4 x 10-6 | |
32463 | NY | 2003 | Chicken | A | - | - | Su | T | Ti | - | K | Ag | - | - | - | - | - | - | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | - | 11 | 12 | 1.1 x 10-5 | |
N1577 | MD | 2004 | Chicken | A | - | - | Su | T | Ti | - | - | Ag | - | - | - | - | - | 1 | 2 | - | 4 | - | 6 | - | - | - | - | 11 | 12 | - | |
N1583 | MD | 2004 | Chicken | A | - | - | Su | T | Ti | - | - | Ag | - | - | - | - | - | 1 | 2 | - | 4 | 5 | - | - | - | - | - | 11 | 12 | - | |
N1591 | MD | 2004 | Chicken | A | - | - | Su | T | Ti | - | K | Ag | - | - | - | - | - | 1 | 2 | - | 4 | - | 6 | 7 | 8 | 9 | - | 11 | 12 | 8.2 x 10-7 | |
N145 | CT | 2004 | Chicken | A | - | - | Su | T | Ti | - | - | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | - | - | - | - | 11 | 12 | 3.8 x 10-5 | |
N167 | CT | 2004 | Chicken | A | - | - | Su | T | Ti | - | - | Ag | - | - | - | - | - | 1 | 2 | - | 4 | - | 6 | - | - | - | - | 11 | 12 | 3.2 x 10-6 | |
N509 | NY | 2004 | Chicken | A | - | - | Su | T | Ti | - | K | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | - | 11 | 12 | 3.7 x 10-5 | |
N520 | NY | 2004 | Chicken | A | - | - | Su | T | Ti | - | - | Ag | - | - | - | - | - | 1 | 2 | - | 4 | - | 6 | 7 | 8 | 9 | - | 11 | 12 | 2.9 x 10-6 | |
N168 | CT | 2004 | Chicken | A | - | - | Su | T | Ti | - | K | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | - | 11 | 12 | - | |
N169 | CT | 2004 | Chicken | A | - | - | Su | T | Ti | - | - | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | - | 11 | 12 | - | |
N176 | CT | 2004 | Chicken | A | C | S | Su | T | Ti | - | - | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | - | - | - | - | 11 | - | - | |
N177 | CT | 2004 | Chicken | A | - | - | Su | T | Ti | - | - | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | - | 11 | 12 | - | |
N499 | NY | 2004 | Chicken | A | - | - | Su | T | Ti | - | K | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | - | 11 | 12 | - | |
N1563 | MD | 2004 | Chicken | A | - | - | Su | T | Ti | - | K | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | - | 11 | 12 | 9.0 x 10-6 | |
N1575 | MD | 2004 | Chicken | A | - | - | Su | T | Ti | - | - | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | - | 11 | 12 | 8.4 x 10-4 | |
N4528 | CT | 2005 | Chicken | A | - | - | Su | T | Ti | - | K | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | - | - | - | 8 | 9 | - | 11 | 12 | - | |
N4546 | CT | 2005 | Chicken | A | - | - | Su | T | Ti | - | - | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | - | - | - | 8 | 9 | - | 11 | 12 | - | |
N7313 | CT | 2005 | Chicken | A | - | - | Su | T | Ti | - | - | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | - | - | - | 8 | 9 | - | 11 | 12 | - | |
N6424 | NY | 2005 | Chicken | A | - | - | Su | T | Ti | - | K | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | - | 8 | 9 | - | 11 | 12 | 2.5 x 10-4 | |
N6426 | NY | 2005 | Chicken | A | - | - | Su | T | Ti | - | - | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | - | - | 12 | - | |
N6430 | NY | 2005 | Chicken | A | - | - | Su | T | Ti | - | K | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | - | 6 | - | 8 | 9 | - | 11 | 12 | - | |
N6437 | NY | 2005 | Chicken | A | - | - | Su | T | Ti | - | - | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | - | 8 | 9 | - | 11 | 12 | 4.3 x 10-3 | |
N5370 | MD | 2005 | Chicken | A | - | - | Su | T | Ti | - | - | Ag | - | - | - | - | - | - | 2 | 3 | 4 | 5 | 6 | - | 8 | 9 | - | 11 | 12 | - | |
N5379 | MD | 2005 | Chicken | A | - | - | Su | T | Ti | - | - | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | - | - | - | - | - | - | 6.3 x 10-5 | |
N5380 | MD | 2005 | Chicken | A | - | - | Su | T | Ti | - | K | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | - | 8 | 9 | - | 11 | 12 | - | |
N5385 | MD | 2005 | Chicken | A | - | - | Su | T | Ti | - | K | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | - | 6 | 7 | 8 | 9 | - | 11 | 12 | 1 x 10-6 | |
22404 | CT | 2002 | Beef | A | - | S | Su | T | Ti | - | - | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | - | - | - | 10 | 11 | 12 | - | |
22697 | MD | 2002 | Turkey | A | C | S | Su | T | Ti | - | - | Ag | - | - | - | - | Sxt | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | - | |
22699 | MD | 2002 | Pork | A | C | S | Su | T | Ti | - | - | Ag | - | - | - | - | Sxt | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | - | |
22707 | MD | 2002 | Beef | A | C | S | Su | T | Ti | - | - | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | - | - | - | 10 | 11 | 12 | - | |
29461 | CA | 2003 | Beef | A | C | S | Su | T | Ti | - | - | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | - | |
29768 | OR | 2003 | Pork | A | C | S | Su | T | Ti | - | - | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | - | |
N1543 | GA | 2004 | Beef | A | C | S | Su | T | Ti | - | - | Ag | Ax | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | - | - | - | 10 | 11 | 12 | - | |
N635 | OR | 2004 | Beef | A | C | S | Su | T | Ti | - | - | Ag | - | - | - | - | Sxt | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 10 | 11 | 12 | - | ||
N175 | CT | 2004 | Chicken | A | C | S | Su | T | Ti | - | - | Ag | - | - | - | - | - | 1 | 2 | - | 4 | - | 6 | - | - | - | - | 11 | - | 8.3 x 10-5 | |
N6427 | NY | 2005 | Chicken | A | - | S | - | T | Ti | - | - | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | - | 11 | 12 | - | |
N4534 | CT | 2005 | Chicken | A | - | - | Su | T | Ti | - | - | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | - | 11 | 12 | - | |
N6312 | MN | 2005 | Turkey | A | - | S | Su | T | Ti | - | K | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | - | 8 | 9 | - | 11 | 12 | 5.2 x 10-5 | |
N5389 | MD | 2005 | Turkey | A | C | S | Su | T | Ti | - | - | Ag | - | - | - | - | Sxt | 1 | 2 | 3 | 4 | 5 | - | - | - | - | 10 | 11 | 12 | - | |
20592 | CT | 2002 | Turkey | A | - | S | Su | T | Ti | - | - | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | - | - | - | - | 10 | 11 | 12 | - | |
21380 | CT | 2002 | Turkey | A | - | S | Su | T | Ti | G | K | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 3.3 x 10-2 | |
N416 | NM | 2004 | Turkey | A | C | S | Su | T | Ti | G | - | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 9 x 10-4 | |
N418 | NM | 2004 | Turkey | A | C | S | Su | T | Ti | G | K | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 4.4 x 10-2 | |
N633 | OR | 2004 | Chicken | A | C | S | Su | T | Ti | - | - | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 6.3 x 10-2 | |
N4504 | CO | 2005 | Turkey | A | - | - | - | - | Ti | - | - | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | - | |
29440 | MN | 2003 | Beef | A | C | S | Su | T | Ti | - | - | Ag | Ax | - | - | - | - | 1 | 2 | 3 | 4 | 5 | - | - | - | - | - | - | - | - | |
23742 | OR | 2003 | Beef | A | C | S | Su | T | Ti | - | - | Ag | - | - | - | - | Sxt | 1 | 2 | 3 | 4 | 5 | 6 | 7 | - | - | 10 | 11 | 12 | - | |
N187 | S. Bredeney | CT | 2004 | Turkey | A | - | - | Su | T | Ti | G | - | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | - | - | - | - | 10 | 11 | 12 | - |
N6328 | MN | 2005 | Turkey | A | - | S | Su | T | Ti | - | - | Ag | Ax | - | - | - | - | 1 | 2 | 3 | 4 | 5 | - | - | - | - | 10 | 11 | 12 | - | |
10688 | IA | 2002 | Turkey | A | C | S | Su | T | Ti | G | K | Ag | - | N | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | - | 8 | 9 | 10 | 11 | 12 | 4.9 x 10-3 | |
10689 | IA | 2002 | Turkey | A | C | S | Su | T | Ti | G | K | Ag | - | N | - | - | - | 1 | - | 3 | - | 5 | - | - | 8 | 9 | 10 | 11 | - | 9.3 x 10-3 | |
10698 | IA | 2002 | Turkey | A | C | S | Su | T | Ti | G | K | Ag | - | - | - | - | - | 1 | - | - | - | 5 | - | - | - | - | 10 | 11 | - | - | |
10978 | IA | 2002 | Turkey | A | C | S | Su | T | Ti | G | K | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 1 x 10-2 | |
11226 | IA | 2002 | Turkey | A | C | S | Su | T | Ti | G | K | Ag | - | - | - | - | - | 1 | - | 3 | - | 5 | - | - | 8 | 9 | 10 | 11 | 12 | 2.5 x 10-3 | |
11228 | IA | 2002 | Turkey | A | C | S | Su | T | Ti | G | K | Ag | - | - | - | - | - | 1 | - | 3 | - | 5 | - | - | 8 | 9 | 10 | 11 | 12 | 1.2 x 10-3 | |
12905 | IA | 2002 | Turkey | A | C | S | Su | T | Ti | G | K | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | - | - | - | - | 10 | 11 | 12 | - | |
12906 | IA | 2002 | Turkey | A | C | S | Su | T | Ti | G | K | Ag | - | - | - | - | - | 1 | - | 3 | - | 5 | - | - | - | - | 10 | 11 | 12 | - | |
13280 | IA | 2002 | Turkey | A | C | S | Su | T | Ti | G | K | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 7.9 x 10-4 | |
3311b | ND | 1997 | Calf | A | C | S | Su | T | Ti | G | K | Ag | Ax | N | Cp | - | - | 1 | - | 3 | - | 5 | - | - | 8 | 9 | 10 | 11 | 12 | 3.6 x 10-4 | |
19811 | MD | 2002 | Chicken | A | C | S | Su | T | Ti | G | K | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | - | 1.2 x 10-2 | |
4420 | GA | 2002 | Chicken | A | - | S | - | - | Ti | G | K | Ag | Ax | N | - | Ak | - | 1 | - | 3 | - | 5 | - | - | 8 | 9 | 10 | 11 | 12 | 4.5 x 10-3 | |
32645 | GA | 2003 | Turkey | A | C | S | Su | T | Ti | G | K | Ag | - | N | - | - | - | 1 | - | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | - | 3.3 x 10-2 | |
32751 | GA | 2003 | Pork | A | C | S | Su | T | Ti | - | K | Ag | - | - | - | - | - | 1 | - | 3 | - | 5 | 6 | - | - | - | 10 | 11 | - | - | |
N820 | OR | 2004 | Beef | A | C | S | Su | T | Ti | G | - | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | - | - | - | - | 10 | 11 | - | - | |
N935 | OR | 2004 | Chicken | A | C | S | Su | T | Ti | - | - | Ag | - | - | - | - | - | 1 | - | 3 | - | 5 | - | - | - | - | 10 | 11 | - | - | |
N1004 | OR | 2004 | Pork | A | C | S | Su | T | Ti | - | - | Ag | - | - | - | - | - | 1 | 2 | 3 | 4 | 5 | - | - | - | 9 | 10 | 11 | - | - | |
N2024 | GA | 2004 | Beef | A | C | S | Su | T | Ti | - | - | Ag | - | - | - | - | - | 1 | 2 | 3 | - | 5 | - | - | - | - | 10 | 11 | - | - | |
N3878 | TN | 2004 | Chicken | A | C | S | Su | T | Ti | G | - | Ag | - | - | - | - | - | 1 | 2 | 3 | - | - | - | 7 | 8 | 9 | - | 11 | - | 7.2 x 10-3 |
amikacin (Ak), amoxicillin/clavulanate (Ag), ampicillin (A), cefoxitin (Cx), ceftiofur (Ti), ceftriaxone (Ax), chloramphenicol (C), ciprofloxacin (Cp), gentamicin (G), kanamycin (K), nalidixic acid (N), tetracycline (T), streptomycin (S), sulfamethoxazole (Su), trimethoprim/sulfamethoxazole (Sxt).
PCR amplicons tested shown in
Estimated efficiency of transfer to Y. ruckeri; expressed as the number of transconjugants per donor; all transconjugants tested positive for the IncA/C replicon.
The discovery of these MDR IncA/C plasmids in evolutionarily distinct pathogens attests to recent genetic exchange, either directly between these bacterial species or through bacterial intermediates, and it suggests that overlap in the ecological niches of these organisms is sufficient to permit past or future plasmid transmission. This is consistent with the high identity (60 kb, 97% nucleotide identity) found between
While identification of the exact bacterial donor or sequence of events leading to the transfer of pIP1202 to
Our findings reveal a potential for the natural acquisition of multidrug resistance in
Plasmids pIP1202 and pSN254 were assembled from whole genome shotgun sequencing of
Plasmid DNA for PCR was prepared using the QIAprep Spin Miniprep kit according to the manufacturer's procedure (Qiagen). PCR was performed using HotStarTaq (Qiagen) with the following cycling protocol: five min enzyme activation at 95°C, 26 cycles of 45 s at 95°C, 45 s at 55°C and 90 s at 72°C followed by a 10 min final extension step at 72°C. The 13 primer sets used were designed to each amplify a distinct plasmid backbone region displayed in
Conjugative mating experiments were performed using a rifampicin resistant derivative of
Primers used in this study
(0.03 MB DOC)
We thank Chris Campbell, Karen Blickenstaff, Lingxia Jiang and Sara Koenig for technical assistance.