Skip to main content
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
letter
. 2004 Jun;48(6):2342–2343. doi: 10.1128/AAC.48.6.2342-2343.2004

Aeromonas hydrophila with Plasmid-Borne Class A Extended-Spectrum β-Lactamase TEM-24 and Three Chromosomal Class B, C, and D β-Lactamases, Isolated from a Patient with Necrotizing Fasciitis

T Fosse 1,2,*, C Giraud-Morin 1,2, I Madinier 1,2, F Mantoux 1,2, J P Lacour 1,2, J P Ortonne 1,2
PMCID: PMC415595  PMID: 15155251

We report a case of necrotizing fasciitis with probable in vivo transfer of a TEM-24 plasmid-borne extended-spectrum β-lactamase (ESBL) gene from Enterobacter aerogenes to Aeromonas hydrophila. The patient was an 87-year-old female with a leg lesion following a trauma. She had a history of rheumatoid polyarthritis treated by 10 mg of prednisone per day, refractory anemia, and chronic venous insufficiency of the lower limbs. Within 5 days, the infection grew worse and the initial amoxicillin-clavulanic acid antibiotic therapy was replaced with ceftriaxone-metronidazole (1 to 1.5 g daily). Surgical debridement revealed extensive necrosis, and 3 days later, the lesion evolved toward typical necrotizing fasciitis (1, 6), leading to a second surgical intervention for above-knee amputation followed by complete healing.

Routine bacteriological procedures revealed (i) Escherichia coli NI-202 susceptible to most β-lactam compounds, (ii) E. aerogenes NI-203 resistant to all β-lactam antibiotics except imipenem, (iii) A. hydrophila NI-204 resistant to ceftazidime, and (iv) A. hydrophila NI-205 susceptible to ceftazidime (Table 1). Pulsed-field electrophoresis confirmed that A. hydrophila NI-204 and NI-205 derived from a single clone. For β-lactamase analysis, the E. aerogenes isolate was grown in brain heart infusion broth with and without cefoxitin or ceftazidime induction (10 μg/ml) at 37°C before analytical isoelectric focusing with crude sonic cell extracts on polyacrylamide gels (2, 4). Two bands of β-lactamase activity were detected with iodine gel with cefazolin (500 μg/ml) as the substrate, which was suggestive of the production of an inducible cephalosporinase (pI 8.8) and an ESBL (pI 6.5). Aeromonas isolates were grown at 30°C with cefoxitin (10 μg/ml), imipenem (1 μg/ml), or tobramycin (1 μg/ml) induction (5). Analytical isoelectric focusing with penicillin and cefazolin as substrates revealed three bands (pI 7, 7.8, and 8.2) probably corresponding to previously described cephalosporinase-, imipenemase-, and oxacillinase-type inducible β-lactamases (5, 11, 12). A. hydrophila NI-204 produced an additional enzyme similar to E. aerogenes NI-203 ESBL (pI 6.5).

TABLE 1.

β-Lactamases, plasmid content, and MICs (μg/ml) of the clinical isolates and transconjugant strains

Parameter or drug(s) E. aerogenes NI-203 A. hydrophila NI-204 A. hydrophila NI-205 A. hydrophila NI-206 E. coli C1a E. coli C1a NI-207
pI 6.5, 8.8 6.5, 7, 7.8, 8.2 7, 7.8, 8.2 6.5, 7, 7.8, 8.2 None 6.5
Plasmid size (kb) 180 180 180 180
PCR TEM + + +
Amoxicillin >256 >256 >256 >256 2 >256
Amoxicillin + clavulanic acid 8 8 8 16 2 2
Ticarcillin >256 >256 >256 >256 8 >256
Ticarcillin + clavulanic acid 64 32 64 16 8 8
Piperacillin 256 4 2 4 2 64
Piperacillin + tazobactam 16 1 1 1 2 2
Cefoxitin >256 16 16 2 2 2
Cefepime 1 0.25 0.06 0.25 ≤0.03 1
Cefepime + clavulanic acid 0.125 ≤0.03 ≤0.03 ≤0.03 ≤0.03 ≤0.03
Cefuroxime >256 2 1 2 1 1
Cefpirome 2 0.5 0.03 0.5 0.03 1
Cefotaxime >32 0.5 0.06 0.5 0.06 1
Ceftazidime >256 32 0.125 32 0.125 256
Ceftazidime + clavulanic acid >4 0.06 0.06 0.06 0.25 0.25
Imipenem 0.5 4 4 8 0.5 0.5
Amikacin 8 8 2 8 0.25 16
Tobramycin 8 16 2 16 0.25 16
Sulfamethoxazole-trimethoprim >32 >32 0.25 >32 0.5 >32
Ciprofloxacin >32 ≤0.03 ≤0.03 ≤0.03 0.125 0.125

Taking into account resistance to ceftazidime, pI determination, and local epidemiology, the ESBL was presumed to be the plasmid-mediated TEM-24 βlactamase (2-4, 7). The plasmid was transferred from E. aerogenes NI-203 to A. hydrophila NI-205 and to E. coli C1a at a high frequency (10−4). Recipient strains (NI-206 and NI-207, Table 1) presented the same acquired resistance pattern. After plasmid extraction and gel electrophoresis, both wild-type strains (E. aerogenes NI-203, A. hydrophila NI-204) and recipient strains (A. hydrophila NI-206, E. coli C1a NI-207) showed a common 180-kb band, as previously characterized with Enterobacteriaceae, Pseudomonas aeruginosa, and recently A. caviae (4, 7-9). The capacity of Aeromonas salmonicida to maintain either or both of the Pseudomonas and Enterobacteriaceae R factors has already been observed (10). PCR amplification with TEM family-specific primers was applied to E. aerogenes NI-203 and A. hydrophila NI-204 and showed a deduced protein sequence with 100% identity to that of TEM-24 (3, 7).

This report demonstrates probable in vivo transfer of ESBL TEM-24 from E. aerogenes to the genus Aeromonas. It was observed in a wild-type strain of A. hydrophila simultaneously producing the class A, B, C, and D β-lactamases.

REFERENCES

  • 1.Angel, M. F., F. Zhang, M. Jones, J. Henderson, and S. W. Chapman. 2002. Necrotizing fasciitis of the upper extremity resulting from a water moccasin bite. South. Med. J. 95:1090-1094. [PubMed] [Google Scholar]
  • 2.Bradford, P. A. 2001. Extended-spectrum β-lactamases in the 21st century: characterization, epidemiology, and detection of this important resistance threat. Clin. Microbiol. Rev. 14:933-951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 3.Chanal, C., M.-C. Poupart, D. Sirot, R. Labia, J. Sirot, and R. Cluzel. 1992. Nucleotide sequences of CAZ-2, CAZ-6, and CAZ-7 β-lactamase genes. Antimicrob. Agents Chemother. 36:1817-1820. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 4.Dumarche, P., C. De Champs, D. Sirot, C. Chanal, R. Bonnet, and J. Sirot. 2002. TEM derivative-producing Enterobacter aerogenes strains: dissemination of a prevalent clone. Antimicrob. Agents Chemother. 46:1128-1131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 5.Fosse, T., C. Giraud-Morin, I. Madinier, and R. Labia. 2003. Sequence analysis and biochemical characterization of chromosomal CAV-1 (Aeromonas caviae), the parental cephalosporinase of plasmid-mediated AmpC ′FOX' cluster. FEMS Microbiol. Lett. 222:93-98. [DOI] [PubMed] [Google Scholar]
  • 6.Furusu, A., N. Yoshizuka, K. Abe, O. Sasaki, K. Miyazaki, M. Miyazaki, Y. Hirakata, Y. Ozono, T. Harada, and S. Kohno. 1997. Aeromonas hydrophila necrotizing fasciitis and gas gangrene in a diabetic patient on hemodialysis. Nephrol. Dial. Transplant. 12:1730-1734. [DOI] [PubMed] [Google Scholar]
  • 7.Giraud-Morin, C., and T. Fosse. 2003. A seven-year survey of Klebsiella pneumoniae producing TEM-24 extended-spectrum β-lactamase in Nice University Hospital (1994-2000). J. Hosp. Infect. 54:25-31. [DOI] [PubMed] [Google Scholar]
  • 8.Marchandin, H., H. Jean-Pierre, C. De Champs, D. Sirot, H. Darbas, P. F. Perigault, and C. Carriere. 2000. Production of a TEM-24 plasmid-mediated extended-spectrum β-lactamase by a clinical isolate of Pseudomonas aeruginosa. Antimicrob. Agents Chemother. 44:213-216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9.Marchandin, H., S. Godreuil, H. Darbas, H. Jean-Pierre, E. Jumas-Bilak, C. Chanal, and R. Bonnet. 2003. Extended-spectrum β-lactamase TEM-24 in an Aeromonas clinical strain: acquisition from the prevalent Enterobacter aerogenes clone in France. Antimicrob. Agents Chemother. 47:3994-3995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10.Olsen, R. H., and C. D. Wright. 1976. Interaction of Pseudomonas and Enterobacteriaceae plasmids in Aeromonas salmonicida. J. Bacteriol. 128:228-234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11.Rasmussen, B. A., and K. Bush. 1997. Carbapenem-hydrolyzing β-lactamases. Antimicrob. Agents Chemother. 41:223-232. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12.Walsh, T. R., L. Hall, A. P. MacGowan, and P. M. Bennett. 1995. A clinical isolate of Aeromonas sobria with three chromosomally mediated inducible β-lactamases: a cephalosporinase, a penicillinase and a third enzyme displaying carbapenemase activity. J. Antimicrob. Chemother. 35:271-279. [DOI] [PubMed] [Google Scholar]

Articles from Antimicrobial Agents and Chemotherapy are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES